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Figure 1 ChIP results obtained with the Diagenode monoclonal antibody directed against HDAC3 ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H3K9me3 (cat. No. SN-146-100) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (cat. No. kch-maglow-016), using sheared chromatin from 10,000 cells. Two different quantities of antibody (3 and 12 μl per ChIP experiment) were analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and for the inactive gene TSH2B. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2 Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K9me3 A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K9me3 (cat. No. SN-146-100) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.
Figure 3 Western blot analysis using the Diagenode monoclonal antibody directed against H3K9me3 Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode monoclonal antibody against H3K9me3 (cat. No. SN-146-100) 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.
WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.
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