Detection of human Smad3, phosphorylated on Ser423/425, by immunoblotting. Samples: Whole cell lysate from HeLa cells left untreated (- TGF-b) or treated with TGF-b (+ TGF-b). Primary antibody: PhosphoDetect™ Anti-Smad3 (pSer433/435) Rabbit pAb (Cat. No. PK1012) (1:1000) preincubated without peptide (- Smad2 or Smad3 peptide), with phosphopeptide corresponding to the analogous phosphorylation sites in Smad2 (+ Smad2 peptide), or with Smad3 phosphopeptide immunogen (+ Smad3 peptide). Detection: chemiluminescence.
Detection of human Smad3, phosphorylated at Ser423/425, by immunocytochemistry. Sample: HeLa cells left untreated (left panel) or treated with TGF-b (right panel). Primary antibody: PhosphoDetect™ Anti-Smad3 (pSer 423/425)/Smad1 (pSer463/465) Rabbit pAb (Cat. No. PK1012) (1:100). Detection: fluorescence (green) with actin filament staining in red.
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