Application
ChIP was performed with HeLa extracts and either control IgG (2 μg) or Histone H3K36me3 (tri-Methy Lys36) antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNA was detected by qPCR with primers targeting to positive controls the coding region of the active GAPDH and ACTB genes, and to negative controls the promoter and a region located 1 kb upstream of the promoter of the GAPDH gene.
Application
ChIP was performed with K562 extracts and either control IgG (1 μg) or Histone H3K36me3 (tri-Methy Lys36) antibody (0.2, 0.5, 1 and 2 μg per ChIP). The precipitated DNA was detected by qPCR with primers targeting to positive controls the coding region of the active GAPDH and ACTB genes, and to negative controls the promoter and a region located 1 kb upstream of the promoter of the GAPDH gene.
Application
Dot blot analysis of peptides containing modifications or unmodified sequences of histone H3 and H4 using H3K36me3 antibody at a dilution of 1:20,000. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane for analysis.
Application
WB analysis of whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H3K36me3 antibodu at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.
Application
ICC/IF analysis of HeLa cells using H3K36me3 antibody (green) and DAPI (blue). Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H3K36me3 antibody at a dilution of 1:500.