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Monoclonal antibody raised in mouse against human p300 (E1A Binding Protein P300) by DNA immunization in which the C-terminal part of the protein was cloned and expressed.
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 *
5 μg/ChIP
Fig 1, 2
* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μg per IP.
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).
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.
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