CST, 9309S, Rb (4H1) Mouse Monoclonal Antibody

Monoclonal Antibody for studying Rb. Validated for WB,WB,IP,IHC,IF,F,ChIP. Available in 3 sizes. Highly specific and rigorously validated in-house, Rb (4H1) Mouse Monoclonal Antibody (CST #9309) is ready to ship. Product Usage Information For optimal ChIP 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: 1:2000 Fluorescent Western: 1:2000 Immunoprecipitation: 1:100 Immunohistochemistry (Paraffin): 1:800 - 1:3200 Immunofluorescence (Immunocytochemistry): 1:1600 Flow Cytometry (Fixed/Permeabilized): 1:3200 Chromatin IP: 1:200 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 # 61121 . Protocol Available protocols: Western Blotting, Fluorescent Western, Immunoprecipitation, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP Specificity / Sensitivity Rb (4H1) Mouse mAb detects endogenous levels of total Rb protein. The antibody does not cross-react with the Rb homologues p107 or p130, or with other proteins. Species Reactivity: Human, Monkey, Bovine, Pig Source / Purification Monoclonal antibody is produced by immunizing animals with a Rb-C fusion protein containing residues 701-928 of human Rb. The epitope corresponds to a region surrounding His890 of human Rb. Background The retinoblastoma tumor suppressor protein Rb regulates cell proliferation by controlling progression through the restriction point within the G1-phase of the cell cycle (1). Rb has three functionally distinct binding domains and interacts with critical regulatory proteins including the E2F family of transcription factors, c-Abl tyrosine kinase, and proteins with a conserved LXCXE motif (2-4). Cell cycle-dependent phosphorylation by a CDK inhibits Rb target binding and allows cell cycle progression (5). Rb inactivation and subsequent cell cycle progression likely requires an initial phosphorylation by cyclin D-CDK4/6 followed by cyclin E-CDK2 phosphorylation (6). Specificity of different CDK/cyclin complexes has been observed (6-8) and cyclin D1 is required for Ser780 phosphorylation (9). Alternate Names exon 17 tumor GOS561 substitution mutation causes premature stop; GOS563 exon 17 substitution mutation causes premature stop; OSRC; p105-Rb; p110-RB1; pp110; PPP1R130; pRb; prepro-retinoblastoma-associated protein; protein phosphatase 1, regulatory subunit 130; Rb; RB transcriptional corepressor 1; RB1; retinoblastoma 1; retinoblastoma suspectibility protein; Retinoblastoma-associated protein Specification REACTIVITY: H Mk B Pg SENSITIVITY: Endogenous MW (kDa): 110 Source/Isotype: Mouse IgG2a