Description General description Anti-Histone H3, Acetylated (1-20), rabbit polyclonal, recognizes the ~17 kDa acetylated histone H3 protein. It is validated for use in Western blotting, immunocytochemistry & immunoprecipitation. Protein A purified rabbit polyclonal antibody. Recognizes the ~17 kDa acetylated histone H3 protein. Recognizes the ~17 kDa acetylated histone H3 protein. Immunogen Human a diacetylated synthetic peptide [ARTKQTARAcKSTGGAcKAPRKQL(C)] corresponding to amino acids 1-20 of human histone H3, conjugated to KLH Application Immunoblotting (10-500 ng/ml, chemiluminescence) Immunocytochemistry (5 µg/ml) Immunoprecipitation (see comments) Physical form In 150 mM NaCl, 100 mM Tris-glycine, pH 7.4. Reconstitution Following initial thaw, aliquot and freeze (-20°C). Packaging 50 μg in Plastic ampoule Please refer to vial label for lot-specific concentration. Warning Toxicity: Standard Handling (A) Analysis Note Positive Control HeLa cells Other Notes Braunstein, M., et al. 1996. Mol. Cell. Biol.16, 4349. Braunstein, M., et al. 1993. Genes Devel.7, 592. Perry, C.A., et al. 1993. Biochemistry32, 13605. Lin, R., et al. 1989. J. Cell. Biol.108, 1577. For immunocytochemistry, HeLa cells were fixed with 95% ethanol/5% acetic acid and treated with 5 mM sodium butyrate, an inhibitor of histone deacetylases, for 24 h. 10-500 ng/ml of antibody detected acetylated histone H3 in acid-extracted proteins from HeLa cells. Using sodium butyrate, an inhibitor of histone deacetylases, enhances detection of acetylated histone H3. Higher concentrations of antibody also detected unacetylated histone H3. Preliminary evidence from an independent library suggests that phosphorylation of Ser10 on histone H3 may disrupt epitope recognition by this antibody. This antibody has also been reported to work for immunoprecipitation. Variables associated with assay conditions will dictate the optimal working dilution.