CST, 8056T, SIN3A (D9D6) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying SIN3A. Validated for WB,IP,IF,ChIP,C&R. Available in 2 sizes. Highly specific and rigorously validated in-house, SIN3A (D9D6) Rabbit Monoclonal Antibody (CST #8056) 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): 1:200 Chromatin IP: 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, CUT&RUN Specificity / Sensitivity SIN3A (D9D6) Rabbit Monoclonal Antibody recognizes endogenous levels of total SIN3A protein. Based on protein sequence, this antibody is not predicted to cross-react with SIN3B protein. Species Reactivity: Human, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu530 of human SIN3A protein. Background SIN3 was originally identified as a negative regulator of transcription in budding yeast (1,2). Since then, three isoforms of the SIN3 proteins have been identified in mammalian cells, as products of two different genes, SIN3A and SIN3B (3,4). Both SIN3A and SIN3B are nuclear proteins that function as scaffolding subunits for the multi-subunit SIN3 transcriptional repressor complex, containing SIN3A or SIN3B, HDAC1, HDAC2, SDS3, RBBP4/RBAP48, RBBP7/RBAP46, SAP30, and SAP18 (3,4). SIN3 proteins contain four paired amphipathic alpha-helix (PAH) motifs that function in the recruitment of the SIN3 complex to target genes by binding a multitude of DNA-binding transcriptional repressor proteins, including Mad1, p53, E2F4, HCF-1, AML1, Elk-1, NRSF, CTCF, ERα, and MeCP2 (3,4). In addition, SIN3 proteins contain an HDAC interaction domain (HID), which mediates binding of HDAC1 and HDAC2 via the SDS3 bridging protein, and a highly conserved region (HCR) at the carboxy terminus, which contributes to repressor protein binding (3,4). RBBP4 and RBBP7 proteins also bind to SDS3 and contribute to nucleosome binding of the complex. The SIN3 complex functions to repress transcription, in part, by deacetylating histones at target gene promoters (3,4). In addition, recent studies have shown that SIN3 is recruited to the coding regions of repressed and active genes, where it deacetylates histones and suppresses spurious transcription by RNA polymerase II (3,5). In addition to histone deacetylase activity, the SIN3 complex associates with histone methyltransferase (ESET), histone demethylase (JARID1A/RBP2), ATP-dependent chromatin remodeling (SWI/SNF), methylcytosine dioxygenase (TET1), and O-GlcNAc transferase (OGT) activities, all of which appear to contribute to the regulation of target genes (5-9). The SIN3 complex is critical for proper regulation of embryonic development, cell growth and proliferation, apoptosis, DNA replication, DNA repair, and DNA methylation (imprinting and X-chromosome inactivation) (3,4). Alternate Names DKFZp434K2235; FLJ90319; Histone deacetylase complex subunit Sin3a; KIAA0700; Paired amphipathic helix protein Sin3a; SIN3 homolog A, transcription regulator; SIN3 homolog A, transcription regulator (yeast); SIN3 transcription regulator family member A; SIN3A; transcriptional co-repressor Sin3A; Transcriptional corepressor Sin3a; transcriptional regulator, SIN3A; WITKOS Specification REACTIVITY: H Mk SENSITIVITY: Endogenous MW (kDa): 145 Source/Isotype: Rabbit IgG