p300 Antibody - ChIP-seq Grade (C15200211)

Lot	001
Concentration	0.9 µg/µl
Species reactivity	Human
Type	Monoclonal

Purity	Protein A purified
Host	Mouse
Precautions	This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Applications	Suggested dilution	References
ChIP/ChIP-seq ^*	5 μg/ChIP	Fig 1, 2

p300 Antibody ChIP Grade

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against p300

ChIP was performed using HeLa cells, the Diagenode monoclonal antibody against p300 (cat. No. C15200211) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010055), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 μg per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. Quantitative PCR was performed with primers for two genomic regions near the ANKRD32 and IRS2 genes, used as positive controls, and for the coding region of the inactive MYOD1 gene and an intergeic region on chromosome 11, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

p300 Antibody for ChIP-seq

p300 Antibody for ChIP-seq assay

p300 Antibody validated in ChIP-seq

Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against p300

ChIP was performed with 5 μg of the Diagenode antibody against p300 (cat. No. C15200211) on sheared chromatin from 4 million HeLa cells as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 3 mb region of chromosome 5 (figure 2A and B) and in two regions surrounding the IRS2 and ANKRD32 positive control genes (figure 2C and D). The position of the amplicon used for ChIP-qPCR is indicated by an arrow.

Target Description

p300 (UniProt/Swiss-Prot entry Q09472) is a histone acetyltransferase that regulates transcription via chromatin remodelling. As such it is important for cell proliferation and differentiation. p300 is able to acetylate all four core histones in nucleosomes. Acetylation of histones is associated with transcriptional activation. p300 also acetylates non-histone proteins such as HDAC1 leading to its inactivation and modulation of transcription. It has also been identified as a co-activator of HIF1A (hypoxiainducible factor 1 alpha), and thus plays a role in the stimulation of hypoxia-induced genes such as VEGF. Defects in the p300 gene are a cause of Rubinstein-Taybi syndrome and may also play a role in epithelial cancer.

Applications

ChIP-seq (ab)
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ChIP-qPCR (ab)
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Documents

Datasheet p300 C15200211 DATASHEET Datasheet description	Download
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