AML1-ETO Antibody - ChIP-seq Grade (sample size)

Lot	A706-002
Concentration	not determined
Species reactivity	Human
Type	Polyclonal
Purity	Whole antiserum
Host	Rabbit
Precautions	This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Applications	Suggested dilution	References
ChIP/ChIP-seq ^*	4 μl/ChIP	Fig 1, 2
ELISA	1:500	Fig 3
Western Blotting	1:1,000	Fig 4

AML1-ETO Antibody ChIP Grade

Figure 1. ChIP results obtained with the Diagenode antibody directed against AML1-ETO
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.

A.ChIP-seq signals on chromosome 3
AML1-ETO Antibody ChIP-seq Grade
B.Genomic region on chromosome 3 surrounding the OGG1 gene
AML1-ETO Antibody for ChIP-seq
C. Genomic region on chromosome 9 surrounding the FUT7 gene

D. Genomic region on chromosome 12 surrounding the NFE2 gene
AML1-ETO Antibody validated in ChIP-seq

Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against AML1-ETO
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.

AML1-ETO Antibody ELISA Validation

Figure 3. Determination of the antibody titer
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.

AML1-ETO Antibody validated in Western Blot

Figure 4. Western blot analysis using the Diagenode antibody directed against AML1-ETO
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.

This antibody specifically recognizes the AML1 (RUNX1) (UniProtKB/Swiss-Prot entry Q01196) - ETO (RUNX1T1) (UniProtKB/ Swiss-Prot entry Q06455) fusion protein that arises due to a translocation between chromosome 8 and 22 (t(8;21)(q22;q22)). This translocation is one of the most frequent karyotypic abnormalities observed in acute myeloid leukaemia. It produces a chimerical gene made up of the 5’-region of AML1and the 3’-region of ETO. The chimerical protein is thought to associate with the nuclear corepressor/histone deacetylase complex to block hematopoietic differentiation.

WB
Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more

ELISA
Enzyme-linked immunosorbent assay. Read more

ChIP-seq (ab)
Read more

ChIP-qPCR (ab)
Read more

Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...	Download
Epigenetic Antibodies Brochure BROCHURE More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...	Download
Datasheet AML1-ETO C15310197 DATASHEET Datasheet description	Download

AML1-ETO antibody SDS US en	Download
AML1-ETO antibody SDS GB en	Download
AML1-ETO antibody SDS ES es	Download
AML1-ETO antibody SDS DE de	Download
AML1-ETO antibody SDS JP ja	Download
AML1-ETO antibody SDS BE nl	Download
AML1-ETO antibody SDS BE fr	Download
AML1-ETO antibody SDS FR fr	Download

How to properly cite our product/service in your work

We strongly recommend using this: AML1-ETO Antibody - ChIP-seq Grade (sample size) (Hologic Diagenode Cat# C15310197-20 Lot# A706-002). Click here to copy to clipboard.

Using our products or services in your publication? Let us know!

RUNX1-ETO Depletion in t(8;21) AML Leads to C/EBPα- and AP-1-Mediated Alterations in Enhancer-Promoter Interaction.
Ptasinska A, Pickin A, Assi SA, Chin PS, Ames L, Avellino R, Gröschel S, Delwel R, Cockerill PN, Osborne CS, Bonifer C
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 maint...

Suppression of RUNX1/ETO oncogenic activity by a small molecule inhibitor of tetramerization
Schanda J. et al.
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 nerv...

The Hematopoietic Transcription Factors RUNX1 and ERG Prevent AML1-ETO Oncogene Overexpression and Onset of the Apoptosis Program in t(8;21) AMLs
Mandoli A. et al.
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 acetyl...

MEIS2 Is an Oncogenic Partner in AML1-ETO-Positive AML
Vegi NM et al.
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-...

Loss of TET2 in hematopoietic cells leads to DNA hypermethylation of active enhancers and induction of leukemogenesis
Rasmussen KD, Jia G, Johansen JV, Pedersen MT, Rapin N, Bagger FO, Porse BT, Bernard OA, Christensen J, Helin K
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...

Immune evasion by oncogenic proteins of acute myeloid leukemia.
Elias S, Yamin R, Golomb L, Tsukerman P, Stanietsky-Kaynan N, Ben-Yehuda D, Mandelboim O
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 ...

ERG and FLI1 binding sites demarcate targets for aberrant epigenetic regulation by AML1-ETO in acute myeloid leukemia.
Martens JH, Mandoli A, Simmer F, Wierenga BJ, Saeed S, Singh AA, Altucci L, Vellenga E, Stunnenberg HG
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 ...

AML1-ETO Antibody - ChIP-seq Grade (sample size)

How to properly cite our product/service in your work

活动