Human/Mouse Histone Deacetylase 8/HDAC8 Antibody

Detection of Human Histone Deacetylase 8/HDAC8 by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse Histone Deacetylase 8/HDAC8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4359) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Histone Deacetylase 8/HDAC8 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Detection of Histone Deacetylase 8/ HDAC8-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Histone Deacetylase 8/HDAC8/DNA complexes were immunoprecipitated using 5 µg Sheep Anti-Human/Mouse Histone Deacetylase 8/HDAC8 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4359) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # [[catalogNumber:BAF016).]]Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # [[catalogNumber:MAG999]]) and DNA was purified using chelating resin solution. TheIFN-gammapromoter was detected by standard PCR.