H3K4un (3C5) Monoclonal Antibody

HeLa cells were stained with H3K4un (3C5) Monoclonal Antibody (Cat. No. bsm-53012M) 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 H3K4un 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.

To test the specificity an ELISA was performed using a serial dilution of H3K4un (3C5) Monoclonal Antibody (Cat. No. bsm-53012M). The wells were coated with peptides containing the unmodified H3K4 region as well as the mono-, di- and trimethylated H3K4. Figure 2 shows a high specificity of the antibody for the unmodified peptide.

ChIP assays were performed using HeLa cells, the monoclonal antibody against H3K4un (Cat. No. bsm-53012M) and optimized PCR primer sets for qPCR. A titration of the antibody consisting of 1, 5 and 10 \u03bcg per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).