HDAC2 (7G5) Monoclonal Antibody

HeLa cells were stained with HDAC2 (7G5) Monoclonal Antibody (bsm-53026M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the HDAC2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

ChIP assays were performed using HeLa cells, HDAC2 (7G5) Monoclonal Antibody (bsm-53026M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2 and 5 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ACTB, EIF4A2 and LMO4 genes, used as positive controls, and for the Sat2 satellite repeat, used as negative control. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ChIP was performed on sheared chromatin from 4 million HeLa cells using 1 \u03bcg of HDAC2 (7G5) Monoclonal Antibody (Cat. No. C15200201) as described above. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the mouse genome using the BWA algorithm. Figure shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 3 (fig 2A and B), and in a two genomic regions surrounding the LMO4 positive control gene and the MAGEC1 gene (figure 2C and D).