CST, 47573T, HP1 alpha (F3K7M) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying HP1 alpha. Validated for WB,WB,IP,IF,IF,ChIP,C&R,C&T. Available in 2 sizes. Highly specific and rigorously validated in-house, HP1 alpha (F3K7M) Rabbit Monoclonal Antibody (CST #47573) is ready to ship. Product Usage Information For optimal ChIP results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 × 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 . The CUT&Tag dilution was determined using CUT&Tag Assay Kit # 77552 . Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:50 Immunofluorescence (Frozen): 1:200 - 1:800 Immunofluorescence (Immunocytochemistry): 1:100 - 1:400 Chromatin IP: 1:50 CUT&RUN: 1:50 CUT&Tag: 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 # 91787 . Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Chromatin IP, CUT&RUN, CUT&Tag Specificity / Sensitivity HP1 alpha (F3K7M) Rabbit Monoclonal Antibody recognizes endogenous levels of total HP1α protein. This antibody does not cross-react with HP1β or HP1γ protein. Non-specific labeling of the epithelia in testis may be observed in fixed frozen mouse tissue by immunofluorescence. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human HP1α 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 Antigen p25; CBX5; chromobox 5; chromobox homolog 5 (HP1 alpha homolog, Drosophila); Chromobox protein homolog 5; epididymis luminal protein 25; HEL25; Heterochromatin protein 1 homolog alpha; heterochromatin protein 1-alpha; HP1; HP1 alpha; HP1 alpha homolog; HP1-ALPHA; HP1A; HP1Hs alpha Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 25 Source/Isotype: Rabbit IgG