WB
Western blot analysis of various lysates using Acetyl-Histone H3-K27 Rabbit pAb at 1:70000 dilution incubated overnight at 4℃. HeLa cells were treated with TSA (1uM) at 37°C for 18 hours, NIH/3T3 cells were treated with TSA (1uM) at 37°C for 18 hours
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit.
Exposure time: 30 s.
DB
Dot-blot analysis of all sorts of peptides using Acetyl-Histone H3-K27 Rabbit pAb at 1:70000 dilution.
IHC
Immunohistochemistry analysis of paraffin-embedded Human spleen tissue using Acetyl-Histone H3-K27 Rabbit pAb at a dilution of 1:300 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Buffer (pH 6.0) prior to IHC staining.
ICC/IF
Immunofluorescence analysis of HeLa cells (treated with TSA) and HeLa cells (untreated) using Acetyl-Histone H3-K27 Rabbit pAb at a dilution of 1:300 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
CHIP
Chromatin immunoprecipitation was performed with 15 μg of cross-linked chromatin from HeLa cells, using 5 μg of Acetyl-Histone H3-K27 Rabbit pAb and Rabbit IgG isotype control. The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.