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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
<p class="text-center"></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Cost-effective (requires less antibody per reaction)</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
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<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'name' => 'RUNX1-ETO Depletion in t(8;21) AML Leads to C/EBPα- and AP-1-Mediated Alterations in Enhancer-Promoter Interaction.',
'authors' => 'Ptasinska A, Pickin A, Assi SA, Chin PS, Ames L, Avellino R, Gröschel S, Delwel R, Cockerill PN, Osborne CS, Bonifer C',
'description' => '<p>Acute myeloid leukemia (AML) is associated with mutations in transcriptional and epigenetic regulator genes impairing myeloid differentiation. The t(8;21)(q22;q22) translocation generates the RUNX1-ETO fusion protein, which interferes with the hematopoietic master regulator RUNX1. We previously showed that the maintenance of t(8;21) AML is dependent on RUNX1-ETO expression. Its depletion causes extensive changes in transcription factor binding, as well as gene expression, and initiates myeloid differentiation. However, how these processes are connected within a gene regulatory network is unclear. To address this question, we performed Promoter-Capture Hi-C assays, with or without RUNX1-ETO depletion and assigned interacting cis-regulatory elements to their respective genes. To construct a RUNX1-ETO-dependent gene regulatory network maintaining AML, we integrated cis-regulatory element interactions with gene expression and transcription factor binding data. This analysis shows that RUNX1-ETO participates in cis-regulatory element interactions. However, differential interactions following RUNX1-ETO depletion are driven by alterations in the binding of RUNX1-ETO-regulated transcription factors.</p>',
'date' => '2019-09-17',
'pmid' => 'http://www.pubmed.gov/31533028',
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'description' => '<p>RUNX1/ETO, the product of the t(8;21) chromosomal translocation, is required for the onset and maintenance of one of the most common forms of acute myeloid leukemia (AML). RUNX1/ETO has a modular structure and, besides the DN A-binding domain (Runt), contains four evolutionary conserved functional domains named nervy homology regions 1-4 (NHR1 to N HR4). The NHR domains serve as docking sites for a variety of different proteins and in addition the N HR2 domain mediates tetramerization through hydrophobic and ionic /polar interactions . Tetramerization is essential for RUNX1/ETO oncogenic activity. Destabilization of the RUNX1/ETO high molecular weight complex abrogates RUNX1/ETO oncogenic activity. Using a structure-based virtual screening, we identified several small molecule inhibitors mimicking the tetramerization hot spot within the NHR2 domain of RUNX1/ETO. One of these compounds, 7.44, was of particular interest as it showed biological activity in vitro and in vivo.</p>',
'date' => '2017-02-02',
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'authors' => 'Mandoli A. et al.',
'description' => '<p>The t(8;21) acute myeloid leukemia (AML)-associated oncoprotein AML1-ETO disrupts normal hematopoietic differentiation. Here, we have investigated its effects on the transcriptome and epigenome in t(8,21) patient cells. AML1-ETO binding was found at promoter regions of active genes with high levels of histone acetylation but also at distal elements characterized by low acetylation levels and binding of the hematopoietic transcription factors LYL1 and LMO2. In contrast, ERG, FLI1, TAL1, and RUNX1 bind at all AML1-ETO-occupied regulatory regions, including those of the AML1-ETO gene itself, suggesting their involvement in regulating AML1-ETO expression levels. While expression of AML1-ETO in myeloid differentiated induced pluripotent stem cells (iPSCs) induces leukemic characteristics, overexpression increases cell death. We find that expression of wild-type transcription factors RUNX1 and ERG in AML is required to prevent this oncogene overexpression. Together our results show that the interplay of the epigenome and transcription factors prevents apoptosis in t(8;21) AML cells.</p>',
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'description' => '<p>Homeobox genes are known to be key factors in leukemogenesis. Although the TALE family homeodomain factor Meis1 has been linked to malignancy, a role for MEIS2 is less clear. Here, we demonstrate that MEIS2 is expressed at high levels in patients with AML1-ETO-positive acute myeloid leukemia and that growth of AML1-ETO-positive leukemia depends on MEIS2 expression. In mice, MEIS2 collaborates with AML1-ETO to induce acute myeloid leukemia. MEIS2 binds strongly to the Runt domain of AML1-ETO, indicating a direct interaction between these transcription factors. High expression of MEIS2 impairs repressive DNA binding of AML1-ETO, inducing increased expression of genes such as the druggable proto-oncogene YES1. Collectively, these data describe a pivotal role for MEIS2 in AML1-ETO-induced leukemia.</p>',
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'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27346355',
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'authors' => 'Rasmussen KD, Jia G, Johansen JV, Pedersen MT, Rapin N, Bagger FO, Porse BT, Bernard OA, Christensen J, Helin K',
'description' => '<p>DNA methylation is tightly regulated throughout mammalian development, and altered DNA methylation patterns are a general hallmark of cancer. The methylcytosine dioxygenase TET2 is frequently mutated in hematological disorders, including acute myeloid leukemia (AML), and has been suggested to protect CG dinucleotide (CpG) islands and promoters from aberrant DNA methylation. In this study, we present a novel <em>Tet2</em>-dependent leukemia mouse model that closely recapitulates gene expression profiles and hallmarks of human AML1-ETO-induced AML. Using this model, we show that the primary effect of <em>Tet2</em> loss in preleukemic hematopoietic cells is progressive and widespread DNA hypermethylation affecting up to 25% of active enhancer elements. In contrast, CpG island and promoter methylation does not change in a <em>Tet2</em>-dependent manner but increases relative to population doublings. We confirmed this specific enhancer hypermethylation phenotype in human AML patients with <em>TET2</em> mutations. Analysis of immediate gene expression changes reveals rapid deregulation of a large number of genes implicated in tumorigenesis, including many down-regulated tumor suppressor genes. Hence, we propose that TET2 prevents leukemic transformation by protecting enhancers from aberrant DNA methylation and that it is the combined silencing of several tumor suppressor genes in <em>TET2</em> mutated hematopoietic cells that contributes to increased stem cell proliferation and leukemogenesis.</p>',
'date' => '2015-04-17',
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'description' => 'PML-RARA and AML1-ETO are important oncogenic fusion proteins that play a central role in transformation to acute myeloid leukemia (AML). Whether these fusion proteins render the tumor cells with immune evasion properties is unknown. Here we show that both oncogenic proteins specifically downregulate the expression of CD48, a ligand of the natural killer (NK) cell activating receptor 2B4, thereby leading to decreased killing by NK cells. We demonstrate that this process is histone deacetylase (HDAC)-dependent, that it is mediated through the downregulation of CD48 messenger RNA, and that treatment with HDAC inhibitors (HDACi) restores the expression of CD48. Furthermore, by using chromatin immuoprecepitation (ChIP) experiments, we show that AML1-ETO directly interacts with CD48. Finally, we show that AML patients who are carrying these specific translocations have low expression of CD48.',
'date' => '2014-03-06',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/24449212',
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'authors' => 'Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG',
'description' => '<p>ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs and identified ERG/FLI1 as proteins that facilitate binding of oncofusion protein complexes. In addition, we demonstrate that ERG and FLI1 bind the RUNX1 promoter and that shRNA mediated silencing of ERG leads to reduced expression of RUNX1 and AML1-ETO, consistent with a role of ERG in transcriptional activation of these proteins. Finally, we identify H3 acetylation as the epigenetic mark preferentially associated with ETS factor binding. This intimate connection between ERG/FLI1 binding and H3 acetylation implies that one of the molecular strategies of oncofusion proteins such as AML1-ETO and PML-RARα involves the targeting of histone deacetylase activities to ERG/FLI1 bound hematopoietic regulatory sites. Together these results highlight the dual importance of ETS factors in t(8;21) leukemogenesis, both as transcriptional regulators of the oncofusion protein itself as well as proteins that facilitate AML1-ETO binding.</p>',
'date' => '2012-09-14',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/22983443',
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<li>Batch-specific data is available on the website</li>
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<div class="small-10 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'authors' => 'Ptasinska A, Pickin A, Assi SA, Chin PS, Ames L, Avellino R, Gröschel S, Delwel R, Cockerill PN, Osborne CS, Bonifer C',
'description' => '<p>Acute myeloid leukemia (AML) is associated with mutations in transcriptional and epigenetic regulator genes impairing myeloid differentiation. The t(8;21)(q22;q22) translocation generates the RUNX1-ETO fusion protein, which interferes with the hematopoietic master regulator RUNX1. We previously showed that the maintenance of t(8;21) AML is dependent on RUNX1-ETO expression. Its depletion causes extensive changes in transcription factor binding, as well as gene expression, and initiates myeloid differentiation. However, how these processes are connected within a gene regulatory network is unclear. To address this question, we performed Promoter-Capture Hi-C assays, with or without RUNX1-ETO depletion and assigned interacting cis-regulatory elements to their respective genes. To construct a RUNX1-ETO-dependent gene regulatory network maintaining AML, we integrated cis-regulatory element interactions with gene expression and transcription factor binding data. This analysis shows that RUNX1-ETO participates in cis-regulatory element interactions. However, differential interactions following RUNX1-ETO depletion are driven by alterations in the binding of RUNX1-ETO-regulated transcription factors.</p>',
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'description' => '<p>RUNX1/ETO, the product of the t(8;21) chromosomal translocation, is required for the onset and maintenance of one of the most common forms of acute myeloid leukemia (AML). RUNX1/ETO has a modular structure and, besides the DN A-binding domain (Runt), contains four evolutionary conserved functional domains named nervy homology regions 1-4 (NHR1 to N HR4). The NHR domains serve as docking sites for a variety of different proteins and in addition the N HR2 domain mediates tetramerization through hydrophobic and ionic /polar interactions . Tetramerization is essential for RUNX1/ETO oncogenic activity. Destabilization of the RUNX1/ETO high molecular weight complex abrogates RUNX1/ETO oncogenic activity. Using a structure-based virtual screening, we identified several small molecule inhibitors mimicking the tetramerization hot spot within the NHR2 domain of RUNX1/ETO. One of these compounds, 7.44, was of particular interest as it showed biological activity in vitro and in vivo.</p>',
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'description' => '<p>The t(8;21) acute myeloid leukemia (AML)-associated oncoprotein AML1-ETO disrupts normal hematopoietic differentiation. Here, we have investigated its effects on the transcriptome and epigenome in t(8,21) patient cells. AML1-ETO binding was found at promoter regions of active genes with high levels of histone acetylation but also at distal elements characterized by low acetylation levels and binding of the hematopoietic transcription factors LYL1 and LMO2. In contrast, ERG, FLI1, TAL1, and RUNX1 bind at all AML1-ETO-occupied regulatory regions, including those of the AML1-ETO gene itself, suggesting their involvement in regulating AML1-ETO expression levels. While expression of AML1-ETO in myeloid differentiated induced pluripotent stem cells (iPSCs) induces leukemic characteristics, overexpression increases cell death. We find that expression of wild-type transcription factors RUNX1 and ERG in AML is required to prevent this oncogene overexpression. Together our results show that the interplay of the epigenome and transcription factors prevents apoptosis in t(8;21) AML cells.</p>',
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'description' => 'PML-RARA and AML1-ETO are important oncogenic fusion proteins that play a central role in transformation to acute myeloid leukemia (AML). Whether these fusion proteins render the tumor cells with immune evasion properties is unknown. Here we show that both oncogenic proteins specifically downregulate the expression of CD48, a ligand of the natural killer (NK) cell activating receptor 2B4, thereby leading to decreased killing by NK cells. We demonstrate that this process is histone deacetylase (HDAC)-dependent, that it is mediated through the downregulation of CD48 messenger RNA, and that treatment with HDAC inhibitors (HDACi) restores the expression of CD48. Furthermore, by using chromatin immuoprecepitation (ChIP) experiments, we show that AML1-ETO directly interacts with CD48. Finally, we show that AML patients who are carrying these specific translocations have low expression of CD48.',
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'description' => '<p>ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs and identified ERG/FLI1 as proteins that facilitate binding of oncofusion protein complexes. In addition, we demonstrate that ERG and FLI1 bind the RUNX1 promoter and that shRNA mediated silencing of ERG leads to reduced expression of RUNX1 and AML1-ETO, consistent with a role of ERG in transcriptional activation of these proteins. Finally, we identify H3 acetylation as the epigenetic mark preferentially associated with ETS factor binding. This intimate connection between ERG/FLI1 binding and H3 acetylation implies that one of the molecular strategies of oncofusion proteins such as AML1-ETO and PML-RARα involves the targeting of histone deacetylase activities to ERG/FLI1 bound hematopoietic regulatory sites. Together these results highlight the dual importance of ETS factors in t(8;21) leukemogenesis, both as transcriptional regulators of the oncofusion protein itself as well as proteins that facilitate AML1-ETO binding.</p>',
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><span>Polyclonal antibody raised in rabbit against the <strong>AML1-ETO</strong> fusion protein using a KLH-conjugated synthetic peptide.</span></p>',
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIP.png" alt="AML1-ETO Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<div class="row">
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong> with rigorous QC and validation</li>
<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Expert technical support</li>
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'name' => 'RUNX1-ETO Depletion in t(8;21) AML Leads to C/EBPα- and AP-1-Mediated Alterations in Enhancer-Promoter Interaction.',
'authors' => 'Ptasinska A, Pickin A, Assi SA, Chin PS, Ames L, Avellino R, Gröschel S, Delwel R, Cockerill PN, Osborne CS, Bonifer C',
'description' => '<p>Acute myeloid leukemia (AML) is associated with mutations in transcriptional and epigenetic regulator genes impairing myeloid differentiation. The t(8;21)(q22;q22) translocation generates the RUNX1-ETO fusion protein, which interferes with the hematopoietic master regulator RUNX1. We previously showed that the maintenance of t(8;21) AML is dependent on RUNX1-ETO expression. Its depletion causes extensive changes in transcription factor binding, as well as gene expression, and initiates myeloid differentiation. However, how these processes are connected within a gene regulatory network is unclear. To address this question, we performed Promoter-Capture Hi-C assays, with or without RUNX1-ETO depletion and assigned interacting cis-regulatory elements to their respective genes. To construct a RUNX1-ETO-dependent gene regulatory network maintaining AML, we integrated cis-regulatory element interactions with gene expression and transcription factor binding data. This analysis shows that RUNX1-ETO participates in cis-regulatory element interactions. However, differential interactions following RUNX1-ETO depletion are driven by alterations in the binding of RUNX1-ETO-regulated transcription factors.</p>',
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'pmid' => 'http://www.pubmed.gov/31533028',
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'name' => 'Suppression of RUNX1/ETO oncogenic activity by a small molecule inhibitor of tetramerization',
'authors' => 'Schanda J. et al.',
'description' => '<p>RUNX1/ETO, the product of the t(8;21) chromosomal translocation, is required for the onset and maintenance of one of the most common forms of acute myeloid leukemia (AML). RUNX1/ETO has a modular structure and, besides the DN A-binding domain (Runt), contains four evolutionary conserved functional domains named nervy homology regions 1-4 (NHR1 to N HR4). The NHR domains serve as docking sites for a variety of different proteins and in addition the N HR2 domain mediates tetramerization through hydrophobic and ionic /polar interactions . Tetramerization is essential for RUNX1/ETO oncogenic activity. Destabilization of the RUNX1/ETO high molecular weight complex abrogates RUNX1/ETO oncogenic activity. Using a structure-based virtual screening, we identified several small molecule inhibitors mimicking the tetramerization hot spot within the NHR2 domain of RUNX1/ETO. One of these compounds, 7.44, was of particular interest as it showed biological activity in vitro and in vivo.</p>',
'date' => '2017-02-02',
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'name' => 'The Hematopoietic Transcription Factors RUNX1 and ERG Prevent AML1-ETO Oncogene Overexpression and Onset of the Apoptosis Program in t(8;21) AMLs',
'authors' => 'Mandoli A. et al.',
'description' => '<p>The t(8;21) acute myeloid leukemia (AML)-associated oncoprotein AML1-ETO disrupts normal hematopoietic differentiation. Here, we have investigated its effects on the transcriptome and epigenome in t(8,21) patient cells. AML1-ETO binding was found at promoter regions of active genes with high levels of histone acetylation but also at distal elements characterized by low acetylation levels and binding of the hematopoietic transcription factors LYL1 and LMO2. In contrast, ERG, FLI1, TAL1, and RUNX1 bind at all AML1-ETO-occupied regulatory regions, including those of the AML1-ETO gene itself, suggesting their involvement in regulating AML1-ETO expression levels. While expression of AML1-ETO in myeloid differentiated induced pluripotent stem cells (iPSCs) induces leukemic characteristics, overexpression increases cell death. We find that expression of wild-type transcription factors RUNX1 and ERG in AML is required to prevent this oncogene overexpression. Together our results show that the interplay of the epigenome and transcription factors prevents apoptosis in t(8;21) AML cells.</p>',
'date' => '2016-11-15',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/27851970',
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'name' => 'MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML',
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'description' => '<p>Homeobox genes are known to be key factors in leukemogenesis. Although the TALE family homeodomain factor Meis1 has been linked to malignancy, a role for MEIS2 is less clear. Here, we demonstrate that MEIS2 is expressed at high levels in patients with AML1-ETO-positive acute myeloid leukemia and that growth of AML1-ETO-positive leukemia depends on MEIS2 expression. In mice, MEIS2 collaborates with AML1-ETO to induce acute myeloid leukemia. MEIS2 binds strongly to the Runt domain of AML1-ETO, indicating a direct interaction between these transcription factors. High expression of MEIS2 impairs repressive DNA binding of AML1-ETO, inducing increased expression of genes such as the druggable proto-oncogene YES1. Collectively, these data describe a pivotal role for MEIS2 in AML1-ETO-induced leukemia.</p>',
'date' => '2016-07-12',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27346355',
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'authors' => 'Rasmussen KD, Jia G, Johansen JV, Pedersen MT, Rapin N, Bagger FO, Porse BT, Bernard OA, Christensen J, Helin K',
'description' => '<p>DNA methylation is tightly regulated throughout mammalian development, and altered DNA methylation patterns are a general hallmark of cancer. The methylcytosine dioxygenase TET2 is frequently mutated in hematological disorders, including acute myeloid leukemia (AML), and has been suggested to protect CG dinucleotide (CpG) islands and promoters from aberrant DNA methylation. In this study, we present a novel <em>Tet2</em>-dependent leukemia mouse model that closely recapitulates gene expression profiles and hallmarks of human AML1-ETO-induced AML. Using this model, we show that the primary effect of <em>Tet2</em> loss in preleukemic hematopoietic cells is progressive and widespread DNA hypermethylation affecting up to 25% of active enhancer elements. In contrast, CpG island and promoter methylation does not change in a <em>Tet2</em>-dependent manner but increases relative to population doublings. We confirmed this specific enhancer hypermethylation phenotype in human AML patients with <em>TET2</em> mutations. Analysis of immediate gene expression changes reveals rapid deregulation of a large number of genes implicated in tumorigenesis, including many down-regulated tumor suppressor genes. Hence, we propose that TET2 prevents leukemic transformation by protecting enhancers from aberrant DNA methylation and that it is the combined silencing of several tumor suppressor genes in <em>TET2</em> mutated hematopoietic cells that contributes to increased stem cell proliferation and leukemogenesis.</p>',
'date' => '2015-04-17',
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'name' => 'Immune evasion by oncogenic proteins of acute myeloid leukemia.',
'authors' => 'Elias S, Yamin R, Golomb L, Tsukerman P, Stanietsky-Kaynan N, Ben-Yehuda D, Mandelboim O',
'description' => 'PML-RARA and AML1-ETO are important oncogenic fusion proteins that play a central role in transformation to acute myeloid leukemia (AML). Whether these fusion proteins render the tumor cells with immune evasion properties is unknown. Here we show that both oncogenic proteins specifically downregulate the expression of CD48, a ligand of the natural killer (NK) cell activating receptor 2B4, thereby leading to decreased killing by NK cells. We demonstrate that this process is histone deacetylase (HDAC)-dependent, that it is mediated through the downregulation of CD48 messenger RNA, and that treatment with HDAC inhibitors (HDACi) restores the expression of CD48. Furthermore, by using chromatin immuoprecepitation (ChIP) experiments, we show that AML1-ETO directly interacts with CD48. Finally, we show that AML patients who are carrying these specific translocations have low expression of CD48.',
'date' => '2014-03-06',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/24449212',
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'name' => 'ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia.',
'authors' => 'Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG',
'description' => '<p>ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs and identified ERG/FLI1 as proteins that facilitate binding of oncofusion protein complexes. In addition, we demonstrate that ERG and FLI1 bind the RUNX1 promoter and that shRNA mediated silencing of ERG leads to reduced expression of RUNX1 and AML1-ETO, consistent with a role of ERG in transcriptional activation of these proteins. Finally, we identify H3 acetylation as the epigenetic mark preferentially associated with ETS factor binding. This intimate connection between ERG/FLI1 binding and H3 acetylation implies that one of the molecular strategies of oncofusion proteins such as AML1-ETO and PML-RARα involves the targeting of histone deacetylase activities to ERG/FLI1 bound hematopoietic regulatory sites. Together these results highlight the dual importance of ETS factors in t(8;21) leukemogenesis, both as transcriptional regulators of the oncofusion protein itself as well as proteins that facilitate AML1-ETO binding.</p>',
'date' => '2012-09-14',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/22983443',
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<p><span>Polyclonal antibody raised in rabbit against the <strong>AML1-ETO</strong> fusion protein using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIP.png" alt="AML1-ETO Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><span>Polyclonal antibody raised in rabbit against the <strong>AML1-ETO</strong> fusion protein using a KLH-conjugated synthetic peptide.</span></p>',
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIP.png" alt="AML1-ETO Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Batch-specific data is available on the website</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<div class="small-2 columns"><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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'name' => 'RUNX1-ETO Depletion in t(8;21) AML Leads to C/EBPα- and AP-1-Mediated Alterations in Enhancer-Promoter Interaction.',
'authors' => 'Ptasinska A, Pickin A, Assi SA, Chin PS, Ames L, Avellino R, Gröschel S, Delwel R, Cockerill PN, Osborne CS, Bonifer C',
'description' => '<p>Acute myeloid leukemia (AML) is associated with mutations in transcriptional and epigenetic regulator genes impairing myeloid differentiation. The t(8;21)(q22;q22) translocation generates the RUNX1-ETO fusion protein, which interferes with the hematopoietic master regulator RUNX1. We previously showed that the maintenance of t(8;21) AML is dependent on RUNX1-ETO expression. Its depletion causes extensive changes in transcription factor binding, as well as gene expression, and initiates myeloid differentiation. However, how these processes are connected within a gene regulatory network is unclear. To address this question, we performed Promoter-Capture Hi-C assays, with or without RUNX1-ETO depletion and assigned interacting cis-regulatory elements to their respective genes. To construct a RUNX1-ETO-dependent gene regulatory network maintaining AML, we integrated cis-regulatory element interactions with gene expression and transcription factor binding data. This analysis shows that RUNX1-ETO participates in cis-regulatory element interactions. However, differential interactions following RUNX1-ETO depletion are driven by alterations in the binding of RUNX1-ETO-regulated transcription factors.</p>',
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'pmid' => 'http://www.pubmed.gov/31533028',
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'name' => 'Suppression of RUNX1/ETO oncogenic activity by a small molecule inhibitor of tetramerization',
'authors' => 'Schanda J. et al.',
'description' => '<p>RUNX1/ETO, the product of the t(8;21) chromosomal translocation, is required for the onset and maintenance of one of the most common forms of acute myeloid leukemia (AML). RUNX1/ETO has a modular structure and, besides the DN A-binding domain (Runt), contains four evolutionary conserved functional domains named nervy homology regions 1-4 (NHR1 to N HR4). The NHR domains serve as docking sites for a variety of different proteins and in addition the N HR2 domain mediates tetramerization through hydrophobic and ionic /polar interactions . Tetramerization is essential for RUNX1/ETO oncogenic activity. Destabilization of the RUNX1/ETO high molecular weight complex abrogates RUNX1/ETO oncogenic activity. Using a structure-based virtual screening, we identified several small molecule inhibitors mimicking the tetramerization hot spot within the NHR2 domain of RUNX1/ETO. One of these compounds, 7.44, was of particular interest as it showed biological activity in vitro and in vivo.</p>',
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'authors' => 'Mandoli A. et al.',
'description' => '<p>The t(8;21) acute myeloid leukemia (AML)-associated oncoprotein AML1-ETO disrupts normal hematopoietic differentiation. Here, we have investigated its effects on the transcriptome and epigenome in t(8,21) patient cells. AML1-ETO binding was found at promoter regions of active genes with high levels of histone acetylation but also at distal elements characterized by low acetylation levels and binding of the hematopoietic transcription factors LYL1 and LMO2. In contrast, ERG, FLI1, TAL1, and RUNX1 bind at all AML1-ETO-occupied regulatory regions, including those of the AML1-ETO gene itself, suggesting their involvement in regulating AML1-ETO expression levels. While expression of AML1-ETO in myeloid differentiated induced pluripotent stem cells (iPSCs) induces leukemic characteristics, overexpression increases cell death. We find that expression of wild-type transcription factors RUNX1 and ERG in AML is required to prevent this oncogene overexpression. Together our results show that the interplay of the epigenome and transcription factors prevents apoptosis in t(8;21) AML cells.</p>',
'date' => '2016-11-15',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/27851970',
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'name' => 'MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML',
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'description' => '<p>Homeobox genes are known to be key factors in leukemogenesis. Although the TALE family homeodomain factor Meis1 has been linked to malignancy, a role for MEIS2 is less clear. Here, we demonstrate that MEIS2 is expressed at high levels in patients with AML1-ETO-positive acute myeloid leukemia and that growth of AML1-ETO-positive leukemia depends on MEIS2 expression. In mice, MEIS2 collaborates with AML1-ETO to induce acute myeloid leukemia. MEIS2 binds strongly to the Runt domain of AML1-ETO, indicating a direct interaction between these transcription factors. High expression of MEIS2 impairs repressive DNA binding of AML1-ETO, inducing increased expression of genes such as the druggable proto-oncogene YES1. Collectively, these data describe a pivotal role for MEIS2 in AML1-ETO-induced leukemia.</p>',
'date' => '2016-07-12',
'pmid' => 'http://www.ncbi.nlm.nih.gov/pubmed/27346355',
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'authors' => 'Rasmussen KD, Jia G, Johansen JV, Pedersen MT, Rapin N, Bagger FO, Porse BT, Bernard OA, Christensen J, Helin K',
'description' => '<p>DNA methylation is tightly regulated throughout mammalian development, and altered DNA methylation patterns are a general hallmark of cancer. The methylcytosine dioxygenase TET2 is frequently mutated in hematological disorders, including acute myeloid leukemia (AML), and has been suggested to protect CG dinucleotide (CpG) islands and promoters from aberrant DNA methylation. In this study, we present a novel <em>Tet2</em>-dependent leukemia mouse model that closely recapitulates gene expression profiles and hallmarks of human AML1-ETO-induced AML. Using this model, we show that the primary effect of <em>Tet2</em> loss in preleukemic hematopoietic cells is progressive and widespread DNA hypermethylation affecting up to 25% of active enhancer elements. In contrast, CpG island and promoter methylation does not change in a <em>Tet2</em>-dependent manner but increases relative to population doublings. We confirmed this specific enhancer hypermethylation phenotype in human AML patients with <em>TET2</em> mutations. Analysis of immediate gene expression changes reveals rapid deregulation of a large number of genes implicated in tumorigenesis, including many down-regulated tumor suppressor genes. Hence, we propose that TET2 prevents leukemic transformation by protecting enhancers from aberrant DNA methylation and that it is the combined silencing of several tumor suppressor genes in <em>TET2</em> mutated hematopoietic cells that contributes to increased stem cell proliferation and leukemogenesis.</p>',
'date' => '2015-04-17',
'pmid' => 'http://genesdev.cshlp.org/content/early/2015/04/15/gad.260174.115',
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'name' => 'Immune evasion by oncogenic proteins of acute myeloid leukemia.',
'authors' => 'Elias S, Yamin R, Golomb L, Tsukerman P, Stanietsky-Kaynan N, Ben-Yehuda D, Mandelboim O',
'description' => 'PML-RARA and AML1-ETO are important oncogenic fusion proteins that play a central role in transformation to acute myeloid leukemia (AML). Whether these fusion proteins render the tumor cells with immune evasion properties is unknown. Here we show that both oncogenic proteins specifically downregulate the expression of CD48, a ligand of the natural killer (NK) cell activating receptor 2B4, thereby leading to decreased killing by NK cells. We demonstrate that this process is histone deacetylase (HDAC)-dependent, that it is mediated through the downregulation of CD48 messenger RNA, and that treatment with HDAC inhibitors (HDACi) restores the expression of CD48. Furthermore, by using chromatin immuoprecepitation (ChIP) experiments, we show that AML1-ETO directly interacts with CD48. Finally, we show that AML patients who are carrying these specific translocations have low expression of CD48.',
'date' => '2014-03-06',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/24449212',
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'name' => 'ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia.',
'authors' => 'Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG',
'description' => '<p>ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs and identified ERG/FLI1 as proteins that facilitate binding of oncofusion protein complexes. In addition, we demonstrate that ERG and FLI1 bind the RUNX1 promoter and that shRNA mediated silencing of ERG leads to reduced expression of RUNX1 and AML1-ETO, consistent with a role of ERG in transcriptional activation of these proteins. Finally, we identify H3 acetylation as the epigenetic mark preferentially associated with ETS factor binding. This intimate connection between ERG/FLI1 binding and H3 acetylation implies that one of the molecular strategies of oncofusion proteins such as AML1-ETO and PML-RARα involves the targeting of histone deacetylase activities to ERG/FLI1 bound hematopoietic regulatory sites. Together these results highlight the dual importance of ETS factors in t(8;21) leukemogenesis, both as transcriptional regulators of the oncofusion protein itself as well as proteins that facilitate AML1-ETO binding.</p>',
'date' => '2012-09-14',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/22983443',
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'description' => '<p><span>Alternative name: <strong>RUNX1-RUNX1T1 <br /></strong></span></p>
<p><span>Polyclonal antibody raised in rabbit against the <strong>AML1-ETO</strong> fusion protein using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIP.png" alt="AML1-ETO Antibody ChIP Grade" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
<p></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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'description' => '<p><span>Alternative name: <strong>RUNX1-RUNX1T1 <br /></strong></span></p>
<p><span>Polyclonal antibody raised in rabbit against the <strong>AML1-ETO</strong> fusion protein using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIP.png" alt="AML1-ETO Antibody ChIP Grade" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO</strong><br />ChIP assays were performed using Kasumi-1 cells, the Diagenode antibody against AML1-ETO (Cat. No. C15310197) and optimized primer pairs for qPCR. Sheared chromatin from 1.25 million cells and 4 μl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the FUT7, NFE2 and OGG1 genes. Figure 1 shows the occupancy, calculated as the ratio + control/background for which the promoter of the H2B gene was used.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><small><strong>A.</strong>ChIP-seq signals on chromosome 3</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-A.png" alt="AML1-ETO Antibody ChIP-seq Grade" /><br /><small><strong>B.</strong>Genomic region on chromosome 3 surrounding the OGG1 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-B.png" alt="AML1-ETO Antibody for ChIP-seq" /><br /><small><strong>C. </strong>Genomic region on chromosome 9 surrounding the FUT7 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-C.png" alt="AML1-ETO Antibody for ChIP-seq assay" /><br /><small><strong>D. </strong>Genomic region on chromosome 12 surrounding the NFE2 gene</small><br /> <img src="https://www.diagenode.com/img/product/antibodies/C15310197_ChIPseq-D.png" alt="AML1-ETO Antibody validated in ChIP-seq" /></p>
<p></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO </strong><br />ChIP was performed as described above. The IP’d DNA of 6 ChIP’s was pooled and analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 32 bp tags were aligned to the human reference genome (hg18) using the ELAND algorithm. Figure 2 shows the results of the complete chromosome 3 and three genomic regions surrounding the OGG1, FUT7 and NFE2 genes, respectively. The position of the PCR amplicon is indicated with an arrow.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197-elisa.png" alt="AML1-ETO Antibody ELISA Validation" /></p>
<p class="text-center"></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer </strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against AML1-ETO (Cat. No. C15310197). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:32,750.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310197_WB.png" alt="AML1-ETO Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO</strong><br />Nuclear extracts of SKNO-1 cells (15 μg) were analysed by Western blot using the Diagenode antibody against AML1-ETO (Cat. No. C15310197) diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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'authors' => 'Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG',
'description' => '<p>ERG and FLI1 are closely related members of the ETS family of transcription factors and have been identified as essential factors for the function and maintenance of normal hematopoietic stem cells. Here, genome-wide analysis revealed that both ERG and FLI1 occupy similar genomic regions as AML1-ETO in t(8;21) AMLs and identified ERG/FLI1 as proteins that facilitate binding of oncofusion protein complexes. In addition, we demonstrate that ERG and FLI1 bind the RUNX1 promoter and that shRNA mediated silencing of ERG leads to reduced expression of RUNX1 and AML1-ETO, consistent with a role of ERG in transcriptional activation of these proteins. Finally, we identify H3 acetylation as the epigenetic mark preferentially associated with ETS factor binding. This intimate connection between ERG/FLI1 binding and H3 acetylation implies that one of the molecular strategies of oncofusion proteins such as AML1-ETO and PML-RARα involves the targeting of histone deacetylase activities to ERG/FLI1 bound hematopoietic regulatory sites. Together these results highlight the dual importance of ETS factors in t(8;21) leukemogenesis, both as transcriptional regulators of the oncofusion protein itself as well as proteins that facilitate AML1-ETO binding.</p>',
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'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/22983443',
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