Histone H3 [Dimethyl Lys36] Antibody (C.15200182)

Chromatin Immunoprecipitation: Histone H3 [Dimethyl Lys36] Antibody (C.15200182) [NBP2-59296] - ChIP assays were performed using human HeLa cells, the antibody against H3K36me2 and optimized PCR primer pairs for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit, using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 ug of antibody per ChIP experiment was analyzed. IgG (1 ug/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 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA: Histone H3 [Dimethyl Lys36] Antibody (C.15200182) [NBP2-59296] - Cross reactivity of the antibody directed against H3K36me2. To test the specificity an ELISA was performed using a serial dilution of the antibody against H3K36me2. 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 shows a high specificity of the antibody for the peptide containing the modification of interest.