EHF Antibody - ChIP-seq Grade

Figure 1. ChIP results obtained with the Diagenode antibody directed against EHF
ChIP assays were performed using T47D cells, the Diagenode antibody against EHF (Cat. No. C15410363) 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 consisting of 1, 2, 5 and 10 µg of antibody per ChIP experiment was analyzed. IgG (2 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers for the promoters of the ITGA2 and OAS3 genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls.

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Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against EHF
ChIP was performed on sheared chromatin from 4,000,000 T47D cells using 2 µg of the Diagenode antibody against EHF (Cat. No. C15410363) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 4000. 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 300 kb region of chromosome 1 (figure 2A and B) and in two genomic regions surrounding the ITGA2 and OAS3 positive control genes, respectively (figure 2C and D).

Figure 3. Determination of the antibody titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Diagenode antibody directed against EHF (Cat. No. C15410363). 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 1:98,000.