CST, 8676S, HP1 beta (D2F2) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying HP1-beta. Validated for WB,IP,IF,ChIP,ChIP,C&R. Available in 2 sizes. Highly specific and rigorously validated in-house, HP1 beta (D2F2) Rabbit Monoclonal Antibody (CST #8676) 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. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . Western Blotting: 1:1000 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:800 Chromatin IP: 1:50 Chromatin IP-seq: 1:50 CUT&RUN: 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, Immunofluorescence (Immunocytochemistry), Chromatin IP, Chromatin IP-seq, CUT&RUN Specificity / Sensitivity HP1 beta (D2F2) Rabbit Monoclonal Antibody recognizes endogenous levels of total HP1β protein. This antibody does not cross-react with other HP1 proteins, including HP1α and HP1γ. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln69 of human HP1β protein. Background Heterochromatin protein 1 (HP1) is a family of heterochromatic adaptor molecules involved in both gene silencing and higher-order chromatin structure (1). All three HP1 family members (α, β, and γ) are primarily associated with centromeric heterochromatin; however, HP1β and γ also localize to euchromatic sites in the genome (2,3). HP1 proteins are approximately 25 kDa and contain a conserved amino-terminal chromodomain, followed by a variable hinge region and a conserved carboxy-terminal chromoshadow domain. The chromodomain facilitates binding to histone H3 tri-methylated at Lys9, a histone "mark" closely associated with centromeric heterochromatin (4,5). The variable hinge region binds both RNA and DNA in a sequence-independent manner (6). The chromoshadow domain mediates the dimerization of HP1 proteins, in addition to binding multiple proteins implicated in gene silencing and heterochromatin formation, including the SUV39H histone methyltransferase, the DNMT1 and DNMT3a DNA methyltransferases, and the p150 subunit of chromatin assembly factor 1 (CAF-1) (7-9). In addition to contributing to heterochromatin formation and propagation, HP1 and SUV39H1 are also found complexed with retinoblastoma (Rb) and E2F6 proteins, both of which function to repress euchromatic gene transcription in quiescent cells (10,11). HP1 proteins are subject to multiple post-translational modifications, including phosphorylation, acetylation, methylation, ubiquitination, and sumoylation, suggesting various means of regulation (12-14). Alternate Names CBX; CBX1; chromobox 1; chromobox homolog 1 (HP1 beta homolog Drosophila ); Chromobox protein homolog 1; Heterochromatin protein 1 homolog beta; heterochromatin protein 1-beta; Heterochromatin protein p25; heterochromatin protein p25 beta; HP1 beta; HP1 beta homolog; HP1-BETA; HP1Hs-beta; HP1Hsbeta; M31; MOD1; Modifier 1 protein; p25beta Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 25 Source/Isotype: Rabbit IgG