CST, 13820L, Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying Smad5 (Ser463/Ser465) phosphate/Smad1 (Ser463/Ser465) phosphate/Smad9 (Ser465/Ser467) phosphate. Validated for WB,IP,IF,F,ChIP. Available in 3 sizes. Highly specific and rigorously validated in-house, Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit Monoclonal Antibody (CST #13820) is ready to ship. Product Usage Information For optimal ChIP 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. Western Blotting: 1:1000 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:400 - 1:1600 Flow Cytometry (Fixed/Permeabilized): 1:200 - 1:800 Chromatin IP: 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), Flow Cytometry (Fixed/Permeabilized), Chromatin IP Specificity / Sensitivity Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit Monoclonal Antibody recognizes endogenous levels of SMAD1 and SMAD5 protein when phosphorylated at Ser463/465 and SMAD9 (SMAD8) protein when phosphorylated at Ser465/467. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human SMAD1 and SMAD5 protein. Background Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β superfamily of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate SMAD1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as SMAD5 and SMAD9 (SMAD8) at their corresponding sites. These phosphorylated SMADs dimerize with the coactivating SMAD4 and translocate to the nucleus, where they regulate the transcription of target genes (5). MAP kinases and CDKs 8 and 9 are also reported to phosphorylate residues in the linker region of SMAD1, including Ser206. Phosphorylation of SMAD1 at Ser206 recruits Smurf1 to the linker region and leads to the degradation of SMAD1 (6). Phosphorylation at this site also promotes SMAD1 transcriptional activity by recruiting YAP to the linker region (7). Alternate Names BSP-1; BSP1; DKFZp781C1895; DKFZp781O1323; Dwfc; hSMAD1; hSmad5; JV4-1; JV41; JV5-1; MAD homolog 1; MAD homolog 5; MAD homolog 9; Mad-related protein 1; MAD, mothers against decapentaplegic homolog 1; MAD, mothers against decapentaplegic homolog 5; MADH1; MADH5; MADH6; MADH9; MADR1; Mothers against decapentaplegic homolog 1; Mothers against decapentaplegic homolog 5; Mothers against decapentaplegic homolog 9; mothers against decapentaplegic, drosophila, homolog of, 5; Mothers against decapentaplegic, drosophila, homolog of, 9; Mothers against DPP homolog 1; Mothers against DPP homolog 5; Mothers against DPP homolog 9; PPH2; SMA- and MAD-related protein 5; SMAD 1; SMAD 5; SMAD 9; SMAD family member 1; SMAD family member 5; SMAD family member 9; SMAD, mothers against DPP homolog 1; SMAD, mothers against DPP homolog 5; SMAD, mothers against DPP homolog 9; SMAD1; SMAD5; SMAD8; SMAD8/9; SMAD8A; SMAD8B; SMAD9; TGF-beta signaling protein 1; transforming growth factor-beta signaling protein 1; Transforming growth factor-beta-signaling protein 1 Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 60 Source/Isotype: Rabbit IgG