WB 1:1000,IHC(P) 1:50-100
|Purification:||Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser423/425 of Smad3.|
Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-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 8, 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-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6–8). Following stimulation by TGF-, Smad2 and Smad3 become phosphorylated at carboxyl terminal serine residues (Ser465 and 467 on Smad2; Ser423 and 425 on Smad3) by TGF-Receptor I. Phosphorylated Smad 2/3 can complex with Smad4 and translocate to the nucleus to regulate gene expression (9-11).Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb detects endogenous levels of Smad3 when phosphorylated at Ser423/425. This antibody does not cross-react with other family members.
|Buffer:||PBS with 0.02% sodium azide, 50% glycerol, pH7.3.|
|Storage:||Store at -20℃. Avoid freeze / thaw cycles.|
tfistern blot analysis of extracts of C6 cells, using Phospho-Smad3-S423/S425 Rabbit mAb (AP0727) at 1:1000 dilution.C6 cells were treated by PMA/TPA (200 nM) at 37℃ for 10 minutes. Secondary antibody: HRP Goat Anti- Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL West Pico Plus Exposure time: 60s.
Immunohistochemistry of paraffinembedded human esophageal using Phospho-Smad3-S423/S425 antibody at dilution of 1:100 (40x lens).
Immunohistochemistry of paraffinembedded mouse testis using Phospho-Smad3-S423/S425 antibody at dilution of 1:100 (40x lens).
Immunohistochemistry of paraffinembedded rat brain using Phospho- Smad3-S423/S425 antibody at dilution of 1:100 (40x lens).
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