Application
ELISA was performed using a serial dilution of Histone H3R17me2K18ac (di-Methyl Arg17, acetyl Lys18) antibody in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:28,800.
Application
ChIP was performed with HeLa extracts and either control IgG (2 μg) or Histone H3R17me2K18ac (di-Methyl Arg17, acetyl Lys18) antibody (1, 2, 5 and 10 μg per ChIP experiment). The precipitated DNAwas detected by qPCR with primers targeting to positive controls the promoters of the active EIF4A2 and c-fos genes, and to negative controls the inactive MYOD1 gene and the Sat2 satellite repeat.
Application
Dot blot analysis of peptides containing histone modifications and the unmodified H3R17K18 using H3R17me2K18ac 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 H3R17me2K18ac antibody at a dilution of 1:1,000 in TBS-Tween containing 5% skimmed milk.
Application
ICC/IF analysis of NIH3T3 cells using H3R17me2K18ac antibody (green) and DAPI (blue). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with H3R17me2K18ac antibody at a dilution of 1:500.