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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K36me2 ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K36me2 (Cat. No. C15200182) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010020), using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a genomic region upstream of the TGM2 gene, used as a positive control, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. 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 of the Diagenode monoclonal antibody directed against H3K36me2 To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K36me2 (Cat. No. C15200182). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure 2 shows a high specificity of the antibody for the peptide containing the modification of interest.
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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