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<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
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<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
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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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<div class="small-12 medium-9 large-9 columns">
<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><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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<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>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-a.png" alt="PPARg Antibody ChIP-seq Grade " /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-b.png" alt="PPARg Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-c.png" alt="PPARg Antibody ChIP-seq assay " /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-d.png" alt="PPARg Antibody validated in ChIP-seq " /></p>
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<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
<div class="small-6 columns">
<p><strong>Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). 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 >1:1,000,000.</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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<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><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-b.png" alt="PPARg Antibody for ChIP-seq" /></p>
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'description' => 'PPARG (UniProtKB/Swiss-Prot entry P37231) is a nuclear hormone receptor which binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Like many other nuclear hormone receptors, PPARG forms a heterodimer with the retinoid X receptor (RXR) leading to transcriptional regulation of various genes including acyl-CoA oxidase and cytochrome P450 A6. PPARG has been implicated in adipocyte differentiation and glucose homeostasis and in various diseases such as obesity, diabetes, atherosclerosis and cancer.',
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'concentration' => '1.9 µg/µl',
'reactivity' => 'Human: positive. Other species: not tested.',
'type' => 'Polyclonal',
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'application_table' => '<table>
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<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:10,000</td>
<td>Fig 3</td>
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<tr>
<td>Western blotting</td>
<td>not recommended</td>
<td></td>
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<p></p>
<p>*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µg per IP.</p>',
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'description' => '<p><strong>Other names: </strong> PPAR-gamma, NR1C3</p>
<p>Polyclonal antibody raised in rabbit against human <strong>PPARg ((peroxisome proliferator-activated receptor gamma),</strong> using two KLH-conjugated synthetic peptides from the central and the N-terminal part of the protein, respectively.</p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-chip.png" alt="PPARg Antibody ChIP Grade" /></center></div>
<div class="small-6 columns">
<p><strong>ChIP results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed with the Diagenode antibody against PPARg (Cat. No. C15410367) on sheared chromatin from 4,000,000 Capan2 cells with the iDeal ChIP-seq kit for TF’s. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the HIST1H4C and LAMP1 genes, used as positive controls, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="row">
<div class="small-12 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-a.png" alt="PPARg Antibody ChIP-seq Grade " /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-b.png" alt="PPARg Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-c.png" alt="PPARg Antibody ChIP-seq assay " /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-d.png" alt="PPARg Antibody validated in ChIP-seq " /></p>
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<div class="row">
<div class="small-12 columns">
<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
<div class="small-6 columns">
<p><strong>Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). 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 >1:1,000,000.</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>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></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>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<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>
<div class="small-12 medium-3 large-3 columns">
<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>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</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>
<p></p>
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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>
<div class="row">
<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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'description' => '<p>Resolution of inflammation is an active process that is tightly regulated to achieve repair and tissue homeostasis. In the absence of resolution, persistent inflammation underlies the pathogenesis of chronic lung disease such as chronic obstructive pulmonary disease (COPD) with recurrent exacerbations. Over the course of inflammation, macrophage programming transitions from pro-inflammatory to pro-resolving, which is in part regulated by the nuclear receptor Peroxisome Proliferator-Activated Receptor γ (PPARγ). Our previous work demonstrated an association between Fatty Acid Binding Protein 5 (FABP5) expression and PPARγ activity in peripheral blood mononuclear cells of healthy and COPD patients. However, a role for FABP5 in macrophage programming has not been examined. Here, using a combination of in vitro and in vivo approaches, we demonstrate that FABP5 is necessary for PPARγ activation. In turn, PPARγ acts directly to increase FABP5 expression in primary human alveolar macrophages. We further illustrate that lack of FABP5 expression promotes a pro-inflammatory macrophage programming with increased secretion of pro-inflammatory cytokines and increased chromatin accessibility for pro-inflammatory transcription factors (e.g., NF-κB and MAPK). And finally, real-time cell metabolic analysis using the Seahorse technology shows an inhibition of oxidative phosphorylation in FABP5-deficient macrophages. Taken together, our data indicate that FABP5 and PPARγ reciprocally regulate each other's expression and function, consistent with a novel positive feedback loop between the two factors that mediates macrophage pro-resolving programming. Our studies highlight the importance of defining targets and regulatory mechanisms that control the resolution of inflammation and may serve to inform novel interventional strategies directed towards COPD.</p>',
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'name' => 'PPARɣ drives IL-33-dependent ILC2 pro-tumoral functions.',
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'description' => '<p>Group 2 innate lymphoid cells (ILC2s) play a critical role in protection against helminths and in diverse inflammatory diseases by responding to soluble factors such as the alarmin IL-33, that is often overexpressed in cancer. Nonetheless, regulatory factors that dictate ILC2 functions remain poorly studied. Here, we show that peroxisome proliferator-activated receptor gamma (PPARγ) is selectively expressed in ILC2s in humans and in mice, acting as a central functional regulator. Pharmacologic inhibition or genetic deletion of PPARγ in ILC2s significantly impair IL-33-induced Type-2 cytokine production and mitochondrial fitness. Further, PPARγ blockade in ILC2s disrupts their pro-tumoral effect induced by IL-33-secreting cancer cells. Lastly, genetic ablation of PPARγ in ILC2s significantly suppresses tumor growth in vivo. Our findings highlight a crucial role for PPARγ in supporting the IL-33 dependent pro-tumorigenic role of ILC2s and suggest that PPARγ can be considered as a druggable pathway in ILC2s to inhibit their effector functions. Hence, PPARγ targeting might be exploited in cancer immunotherapy and in other ILC2-driven mediated disorders, such as asthma and allergy.</p>',
'date' => '2021-05-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33953160',
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'name' => 'PPARg polyclonal antibody datasheet',
'description' => '<p>Polyclonal antibody raised in rabbit against human HDAC3 (Histone deacetylase 3), using two KLH-conjugated synthetic peptides from the central and the N-terminal part of the protein, respectively.</p>',
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'description' => '<p>Group 2 innate lymphoid cells (ILC2s) play a critical role in protection against helminths and in diverse inflammatory diseases by responding to soluble factors such as the alarmin IL-33, that is often overexpressed in cancer. Nonetheless, regulatory factors that dictate ILC2 functions remain poorly studied. Here, we show that peroxisome proliferator-activated receptor gamma (PPARγ) is selectively expressed in ILC2s in humans and in mice, acting as a central functional regulator. Pharmacologic inhibition or genetic deletion of PPARγ in ILC2s significantly impair IL-33-induced Type-2 cytokine production and mitochondrial fitness. Further, PPARγ blockade in ILC2s disrupts their pro-tumoral effect induced by IL-33-secreting cancer cells. Lastly, genetic ablation of PPARγ in ILC2s significantly suppresses tumor growth in vivo. Our findings highlight a crucial role for PPARγ in supporting the IL-33 dependent pro-tumorigenic role of ILC2s and suggest that PPARγ can be considered as a druggable pathway in ILC2s to inhibit their effector functions. Hence, PPARγ targeting might be exploited in cancer immunotherapy and in other ILC2-driven mediated disorders, such as asthma and allergy.</p>',
'date' => '2021-05-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33953160',
'doi' => '10.1038/s41467-021-22764-2',
'modified' => '2021-12-21 16:47:06',
'created' => '2021-12-06 15:53:19',
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$externalLink = ' <a href="https://www.ncbi.nlm.nih.gov/pubmed/33953160" target="_blank"><i class="fa fa-external-link"></i></a>'include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
Notice (8): Undefined variable: campaign_id [APP/View/Products/view.ctp, line 755]Code Context<!-- BEGIN: REQUEST_FORM MODAL -->
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'name' => 'PPARg polyclonal antibody',
'description' => '<p><strong>Other names: </strong> PPAR-gamma, NR1C3</p>
<p>Polyclonal antibody raised in rabbit against human HDAC3 (Histone deacetylase 3), using two KLH-conjugated synthetic peptides from the central and the N-terminal part of the protein, respectively.</p>
<script src="chrome-extension://hhojmcideegachlhfgfdhailpfhgknjm/web_accessible_resources/index.js"></script>',
'label1' => 'Validation data',
'info1' => '<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-chip.png" alt="PPARg Antibody ChIP Grade" /></center></div>
<div class="small-6 columns">
<p><strong>ChIP results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed with the Diagenode antibody against PPARg (Cat. No. C15410367) on sheared chromatin from 4,000,000 Capan2 cells with the iDeal ChIP-seq kit for TF’s. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the HIST1H4C and LAMP1 genes, used as positive controls, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-a.png" alt="PPARg Antibody ChIP-seq Grade " /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-b.png" alt="PPARg Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-c.png" alt="PPARg Antibody ChIP-seq assay " /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410367-chipseq-d.png" alt="PPARg Antibody validated in ChIP-seq " /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
<div class="small-6 columns">
<p><strong>Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). 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 >1:1,000,000.</p>
</div>
</div>',
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'info2' => '<p>PPARG (UniProtKB/Swiss-Prot entry P37231) is a nuclear hormone receptor which binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Like many other nuclear hormone receptors, PPARG forms a heterodimer with the retinoid X receptor (RXR) leading to transcriptional regulation of various genes including acyl-CoA oxidase and cytochrome P450 A6. PPARG has been implicated in adipocyte differentiation and glucose homeostasis and in various diseases such as obesity, diabetes, atherosclerosis and cancer.</p>
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'description' => 'PPARG (UniProtKB/Swiss-Prot entry P37231) is a nuclear hormone receptor which binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Like many other nuclear hormone receptors, PPARG forms a heterodimer with the retinoid X receptor (RXR) leading to transcriptional regulation of various genes including acyl-CoA oxidase and cytochrome P450 A6. PPARG has been implicated in adipocyte differentiation and glucose homeostasis and in various diseases such as obesity, diabetes, atherosclerosis and cancer.',
'clonality' => '',
'isotype' => '',
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'classification' => '',
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<thead>
<tr>
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<th>References</th>
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<tr>
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<td>5 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:10,000</td>
<td>Fig 3</td>
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<tr>
<td>Western blotting</td>
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<p>*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µg per IP.</p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
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'slug' => 'pparg-polyclonal-antibody',
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<p><strong>ChIP results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed with the Diagenode antibody against PPARg (Cat. No. C15410367) on sheared chromatin from 4,000,000 Capan2 cells with the iDeal ChIP-seq kit for TF’s. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the HIST1H4C and LAMP1 genes, used as positive controls, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
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<p><strong>Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). 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 >1:1,000,000.</p>
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<p><strong>ChIP-seq results obtained with the Diagenode antibody directed against PPARg</strong><br />ChIP was performed on sheared chromatin from 4,000,000 Capan2 cells using 5 µg of the Diagenode antibody against PPARg (Cat. No. C15410367) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 1.5 Mb region of chromosome 1 (figure 2A and B), in a genomic region of chromosome 6 containing several HIST1 genes (figure 2C), and in a genomic region of the X-chromosome surrounding the LAMP1 positive control gene (figure 2D).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15410367-elisa.png" alt="PPARg Antibody ELISA Validation" /></center></div>
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<p><strong>Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against PPARg (Cat. No. C15410367). 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 >1:1,000,000.</p>
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<div class="small-12 medium-9 large-9 columns">
<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>
<div class="small-12 medium-3 large-3 columns">
<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>
<li>Expert technical support</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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<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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