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> H3K4me3 (2G10) Monoclonal Antibody

H3K4me3 (2G10) Monoclonal Antibody

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Cat# bsm-53015M

Size : 50ug

Brand : Bioss

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Phone : +1 850 650 7790

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) 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 H3K4me3 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.
HeLa cells were stained with H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) 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 H3K4me3 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 H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
To test the specificity an ELISA was performed using a serial dilution of H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using human HeLa cells, \r\nH3K4me3 (2G10) Monoclonal Antibody (bsm-53015M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of GAPDH and c-fos genes (positive controls), and exon 2 of the inactive myoglobin (MB) gene and Sat2 satellite repeat (negative controls). Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.
ChIP assays were performed using human HeLa cells, \r\nH3K4me3 (2G10) Monoclonal Antibody (bsm-53015M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of GAPDH and c-fos genes (positive controls), and exon 2 of the inactive myoglobin (MB) gene and Sat2 satellite repeat (negative controls). Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.