CST, 85057T, HDAC3 (D2O1K) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying HDAC3. Validated for WB,IP,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, HDAC3 (D2O1K) Rabbit Monoclonal Antibody (CST #85057) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. Western Blotting: 1:1000 Immunoprecipitation: 1:100 Chromatin IP: 1:50 Chromatin IP-seq: 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Chromatin IP, Chromatin IP-seq Specificity / Sensitivity HDAC3 (D2O1K) Rabbit Monoclonal Antibody recognizes endogenous levels of total HDAC3 protein. This antibody does not cross-react with other HDAC proteins. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly416 of human HDAC3 protein. Background Acetylation of the histone tail causes chromatin to adopt an "open" conformation, allowing increased accessibility of transcription factors to DNA. The identification of histone acetyltransferases (HATs) and their large multiprotein complexes has yielded important insights into how these enzymes regulate transcription (1,2). HAT complexes interact with sequence-specific activator proteins to target specific genes. In addition to histones, HATs can acetylate nonhistone proteins, suggesting multiple roles for these enzymes (3). In contrast, histone deacetylation promotes a "closed" chromatin conformation and typically leads to repression of gene activity (4). Mammalian histone deacetylases can be divided into three classes on the basis of their similarity to various yeast deacetylases (5). Class I proteins (HDACs 1, 2, 3, and 8) are related to the yeast Rpd3-like proteins, those in class II (HDACs 4, 5, 6, 7, 9, and 10) are related to yeast Hda1-like proteins, and class III proteins are related to the yeast protein Sir2. Inhibitors of HDAC activity are now being explored as potential therapeutic cancer agents (6,7). HDAC3 is a nuclear and cytoplasmic protein that deacetylates both histone (H2A, H3, H4) and non-histone substrates (RelA, SRY, p53, MEF2, PCAF and p300/CBP) (8). HDAC3 deacetylase activity is stimulated by interactions with the N-CoR and SMRT co-repressor proteins. Together, these three proteins form a functional complex that represses transcription associated with nuclear hormone receptors and other transcription factors, including Rev-Erb, COUP-TF, DAX1, MAD and Pit-1 (8,9). Phosphorylation of HDAC3 on Ser424 by casein kinase 2 (CK2) also increases HDAC3 deacetylase activity (9). Subsequently, de-phosphorylation by protein phosphatase 4 (PP4) decreases HDAC3 activity (9). Alternate Names HD3; HDAC3; Histone deacetylase 3; KDAC3; RPD3; RPD3-2; SMAP45 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 49 Source/Isotype: Rabbit IgG