CST, 83375S, BRD4 (pan) (E1Y1P) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying BRD4 isoform B/BRD4 isoform A/BRD4 isoform C. Validated for WB,IP,ChIP,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, BRD4 (pan) (E1Y1P) Rabbit Monoclonal Antibody (CST #83375) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 5 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: 1:100 Chromatin IP-seq: 1:100 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 BRD4 (pan) (E1Y1P) Rabbit Monoclonal Antibody recognizes endogenous levels of total BRD4 protein. This antibody detects endogenous BRD4 IsoA and BRD4 IsoC, and BRD4 IsoB when overexpressed. This antibody also detects bands of unknown origin at around 78 and 76 kDa. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro53 of human BRD4 protein. Background Bromodomain-containing protein 4 (BRD4) is a member of the bromodomains and extra terminal (BET) family of proteins, which also includes BRD2, BRD3, and BRDT (1-3). BET family proteins contain two tandem bromodomains and an extra terminal (ET) domain, and bind acetyl lysine residues (3). BRD4 is a chromatin-binding protein with a preference for Lys14 on histone H3 as well as Lys5 and Lys12 on histone H4 (4). BRD4 chromatin binding occurs throughout the cell cycle, including condensed mitotic chromosomes, when the majority of genes are silenced (5). BRD4 association with chromatin during mitosis is thought to be an important part of the bookmarking mechanism to accelerate reactivation of the silenced genes upon exit from mitosis (2,6). BRD4 has been shown to facilitate transcription by recruiting the positive transcription elongation factor b (pTEFb) complex that phosphorylates Ser2 of the heptapeptide repeat of the carboxy-terminal domain of RNA polymerase II, promoting transcription elongation (3,7,8). In addition, BRD4 has been found to be part of the super elongation complex and the polymerase associated factor complex (PAFc) in MLL-fusion derived leukemia cell lines, demonstrating a role for BRD4 in the regulation of transcription elongation (9). Research studies have shown that BRD4 (and BET family proteins) may be promising therapeutic targets for various Myc-driven cancers, such as Burkitt's lymphoma and certain acute myeloid leukemias (1,10,11). Investigators have found molecular inhibition of BET proteins to be effective in inducing apoptosis in various MLL-fusion driven leukemic cell lines by competing BRD3 and BRD4 from chromatin, leading to reduced expression of Bcl-2, Myc, and CDK6 (9). BET inhibition has also been shown to have antitumor activities against nuclear protein in testis (NUT) midline carcinoma cell lines and xenografts in mice where BRD4 is found to be a frequent translocation partner of the NUT protein (12). In addition, BRD4 regulates the expression of some inflammatory genes, and inhibition of BRD4 (and BET family proteins) chromatin binding causes reduced expression of a subset of inflammatory genes in macrophages, leading to protection against endotoxic shock and sepsis (13). Alternate Names BRD4; BRD4 iso2; BRD4 iso3; BRD4 Isoform B; BRD4 Isoform C; Brd4S; bromodomain containing 4; bromodomain-containing 4; Bromodomain-containing protein 4; CAP; chromosome-associated protein; hCG_2000917; HUNK1; HUNKI; MCAP; mitotic chromosome-associated protein; Protein HUNK1 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 200, 110 Source/Isotype: Rabbit IgG