Description General description Purified rabbit monoclonal antibody generated by immunizing rabbits with the specified immunogen and fusing splenocytes with an immortal cell line. Recognizes the ~52 kDa Smad3 protein phosphorylated at Ser423/425. Recognizes the ~52 kDa Smad3 protein phosphorylated at Ser423/425 in HT-1080 cells treated with TGF-β1. This PhosphoDetect Anti-Smad3 (pSer423/425) Rabbit mAb (C25A9) is validated for use in Immunoblotting, Immunoprecipitation for the detection of Smad3 (pSer423/425). Immunogen a synthetic phosphopeptide corresponding to amino acids surrounding the Ser 423/425 phosphorylation sites of Smad3, coupled to KLH. Application Immunoblotting (1:1000) Immunoprecipitation (1:50) Physical form In 150 mM NaCl, 10 mM HEPES, 100 µg/ml BSA, 50% glycerol pH 7.5. Legal Information TWEEN is a registered trademark of Croda International PLC Packaging 50 μL in Plastic ampoule Warning Toxicity: Standard Handling (A) Other Notes Attisano, L., et al. 2002. Science 296, 1646. Moustakas, A., et al. 2001. J. Cell Sci. 114, 4359. Wrana, J. 2000. Science 23, 1. Does not cross-react with other Smad proteins. For immunoblotting incubate the membrane with antibody diluted with TBS containing 5% BSA, 0.1% Tween®-20 detergent overnight at 4°C with gentle shaking. Please note that this is a rabbit monoclonal antibody and requires the use of an anti-rabbit secondary antibody. The immunogen sequence is 100% conserved in mouse, rat, monkey, bovine, xenopus, and zebrafish Smad3, but cross-reactivity has not been tested. Antibody should be titrated for optimal results in individual systems.