Rat anti-Human Histone H3K9ac Monoclonal Antibody (Clone KT163)

Cross reactivity test: Microtiter wells were coated with various peptides. KT163 was used as the primary antibody and peroxidase conjugated goat anti-rat IgG was used as the the secondary antibody. Result: KT163 specifically reacts with acetylated H3K9ac.

Immunohistochemistry: IHC-P analysis in hepatocellular carcinoma tissue by H3K9ac antibody (KT163). Immunohistochemistry was performed using sections of the formalin-fixed paraffin- embedded hepatocellular carcinoma tissue.

Immunoprecipitation: Immunoprecipitation was performed by incubation of 2.5 ug KT163 with HeLa lysate containing 200 ug total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. H3K9ac (KT163) at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-rat IgG was used as the secondary antibody. The isotype control antibody was KT82. Lane 1: 15 ug of HeLa lysate Lane 2: H3K9ac immunoprecipitated from HeLa lysate by KT163 Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody Result: KT163 can immunoprecipitate H3K9ac.

Western Blot: Lysate of MCF-7 treated with various concentrations of sodium butyrate was run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. KT163 at 1 ug/mL was used as the primary antibody and peroxidase conjugated goat anti-rat IgG was used as the secondary antibody. H3K9ac band was visualized using ECL Western Blotting Substrate. Lane 1: 15 ug of MCF-7 lysate Lane 2: 15 ug of MCF-7 lysate treated with 1 mM sodium butyrate for 6 hs Lane 3: 15 ug of MCF-7 lysate treated with 5 mM sodium butyrate for 6 hs Lane 4: 15 ug of MCF-7 lysate treated with 10 mM sodium butyrate for 6 hs Result: KT163 can detect H3K9ac by Western blotting.