CST, 7627T, Acetyl-Histone H3 (Lys14) (D4B9) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying H3 (Lys14) acetylate. Validated for WB,WB,IP,IF,F,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, Acetyl-Histone H3 (Lys14) (D4B9) Rabbit Monoclonal Antibody (CST #7627) 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 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:1600 - 1:6400 Flow Cytometry (Fixed/Permeabilized): 1:1600 - 1:6400 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. For a carrier free (BSA and azide free) version of this product see product # 80697 . Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq Specificity / Sensitivity Acetyl-Histone H3 (Lys14) (D4B9) Rabbit Monoclonal Antibody recognizes endogenous levels of Histone H3 protein only when acetylated at Lys14. This antibody does not cross react with histone H3 acetylated at Lys9, 18, 27, or 56. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding acetylated Lys14 of human Histone H3 protein. Background The nucleosome, made up of four core histone proteins (H2A, H2B, H3, and H4), is the primary building block of chromatin. Originally thought to function as a static scaffold for DNA packaging, histones have now been shown to be dynamic proteins, undergoing multiple types of post-translational modifications, including acetylation, phosphorylation, methylation, and ubiquitination (1,2). Histone acetylation occurs mainly on the amino-terminal tail domains of histones H2A (Lys5), H2B (Lys5, 12, 15, and 20), H3 (Lys9, 14, 18, 23, 27, 36, and 56), and H4 (Lys5, 8, 12, and 16) and is important for the regulation of histone deposition, transcriptional activation, DNA replication, recombination, and DNA repair (1-3). Hyper-acetylation of the histone tails neutralizes the positive charge of these domains and is believed to weaken histone-DNA and nucleosome-nucleosome interactions, thereby destabilizing chromatin structure and increasing the accessibility of DNA to various DNA-binding proteins (4,5). In addition, acetylation of specific lysine residues creates docking sites for a protein module called the bromodomain, which binds to acetylated lysine residues (6). Many transcription and chromatin regulatory proteins contain bromodomains and may be recruited to gene promoters, in part, through binding of acetylated histone tails. Histone acetylation is mediated by histone acetyltransferases (HATs), such as CBP/p300, GCN5L2, PCAF, and Tip60, which are recruited to genes by DNA-bound protein factors to facilitate transcriptional activation (3). Deacetylation, which is mediated by histone deacetylases (HDAC and sirtuin proteins), reverses the effects of acetylation and generally facilitates transcriptional repression (7,8). Alternate Names H3; H3 clustered histone 1; H3 histone family, member A; H3/A; H31; H3C1; H3C10; H3C11; H3C12; H3C2; H3C3; H3C4; H3C6; H3C7; H3C8; H3FA; H3FB; H3FC; H3FC HIST1H3C; H3FD; H3FF; H3FH; H3FI; H3FJ; H3FK; H3FL; HIST1H3A; HIST1H3B; HIST1H3C; HIST1H3D; HIST1H3E; HIST1H3F; HIST1H3G; HIST1H3H; HIST1H3I; HIST1H3J; histone 1, H3a; histone cluster 1 H3 family member a; histone cluster 1, H3a; Histone H3; Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/i; Histone H3/j; Histone H3/k; Histone H3/l Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 17 Source/Isotype: Rabbit IgG