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Figure 1. ChIP results obtained with the Diagenode antibody directed against SAP30 ChIP assays were performed using HeLa cells, the Diagenode antibody against SAP30 (Cat. No. C15410036) and optimized 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 and 5 μg per ChIP experiment was analysed. IgG (1 μg/ IP) was used as negative IP control. QPCR was performed with primers for the EIF2S3 and BRCA1 promoters, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, 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 antibody directed against SAP30 ChIP was performed on sheared chromatin from 4 million HeLa cells as described above using 5 μg of the Diagenode antibody against SAP30 (Cat. No. C15410036). The IP’d DNA was subsequently analysed on an Illumina HiSeq. 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 enrichment along the complete sequence and a 1.5 Mb region of human chromosome 1 (fig 2A and B) and in two genomic regions surrounding the BRCA1 and EIF2S3 genes on chromosome 17 and X, respectively (fig 2C and D).
Figure 3. Western blot analysis using the Diagenode antibody directed against SAP30 Western blot was performed on nuclear extracts from HeLa cells (20 μg) using the Diagenode antibody against SAP30 (Cat. No. C15410036) diluted 1:1000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest is indicated on the right.
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.
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