CST, 9520T, Phospho-SMAD3 (Ser423/425) (C25A9) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying Smad3 (Ser423/Ser425) phosphate. Validated for WB,WB,IP,ChIP. Available in 3 sizes. Highly specific and rigorously validated in-house, Phospho-SMAD3 (Ser423/425) (C25A9) Rabbit Monoclonal Antibody (CST #9520) 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: 1:1000 Simple Western™: 1:250 Immunoprecipitation: 1:50 Chromatin IP: 1:100 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, Chromatin IP Specificity / Sensitivity Phospho-SMAD3 (Ser423/425) (C25A9) Rabbit Monoclonal Antibody detects endogenous levels of SMAD3 when phosphorylated at Ser423/425. This antibody does not cross-react with other family members. Species Reactivity: Human, Mouse, Rat Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser423/425 of human SMAD3. Background Members of the SMAD family of signal transduction molecules are components of a critical intracellular pathway that transmit TGF-β signals from the cell surface into the nucleus. Three distinct classes of SMADs have been defined: the receptor-regulated SMADs (R-SMADs), which include SMAD1, 2, 3, 5, and 9; the common-mediator SMAD (co-SMAD), SMAD4; and the antagonistic or inhibitory SMADs (I-SMADs), SMAD6 and 7 (1-5). Activated type I receptors associate with specific R-SMADs and phosphorylate them on a conserved carboxy-terminal SSXS motif. The phosphorylated R-SMADs dissociate from the receptor and form a heteromeric complex with SMAD4, initiating translocation of the heteromeric SMAD complex to the nucleus. Once in the nucleus, SMADs recruit a variety of DNA binding proteins that function to regulate transcriptional activity (6-8). Following stimulation by TGF-β, Smad2 and Smad3 become phosphorylated at carboxyl terminal serine residues (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-β Receptor I. Phosphorylated Smad 2/3 can complex with Smad4 and translocate to the nucleus to regulate gene expression (9-11). Alternate Names DKFZp586N0721; DKFZp686J10186; hMAD-3; hSMAD3; HSPC193; HsT17436; JV15-2; LDS1C; LDS3; MAD homolog 3; mad homolog JV15-2; mad protein homolog; MAD, mothers against decapentaplegic homolog 3; Mad3; MADH3; MGC60396; Mothers against decapentaplegic homolog 3; Mothers against DPP homolog 3; SMA- and MAD-related protein 3; SMAD 3; SMAD family member 3; SMAD, mothers against DPP homolog 3; SMAD3 Specification REACTIVITY: H M R SENSITIVITY: Endogenous MW (kDa): 52 Source/Isotype: Rabbit IgG