H3K4me2 (4B4) Monoclonal Antibody

HeLa cells were stained with H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M) 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 labelled with the H3K4me2 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 staining is shown on the right.

To test the specificity an ELISA was performed using a serial dilution of the H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the dimethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.

ChIP assays were performed using HeLa cells, H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 5 and 10 \u03bcg per ChIP was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).