Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
Specificity
This affinity-purified antibody is directed against the phosphorylated form of Human Smad3 protein at the pSer423 and pSer425 residues. The product was affinity purified from monospecific antiserum by Immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against Human Smad3 pSer423 and pSer425 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human Smad3 is minimal by ELISA and western blot. Expect reactivity against phosphorylated Smad1 and Smad5. Negligible reactivity is seen against other phosphorylated Smad family members.
Purification
Immunoaffinity Chromatography.
Immunogen
Synthetic peptide corresponding to amino acids 417-425 of Human SMAD3 protein
SMAD3
Reactivity: Human, Mouse, Rat
WB, IHC (p), FACS, IF (cc)
Host: Rabbit
Monoclonal
3D1
unconjugated
Application Notes
Suitable for use in ELISA, Immunohistochemistry and Western blot. Expect a bandapproximately 48 kDa in size corresponding to phosphorylated Smad3 protein by Westernblotting in the appropriate stimulated tissue or cell lysate or extract. Less than 0.2%reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for dual phosphorylated pS423 and pS425 of Smad3. Stimulation with 2 ng/ml TGF-beta for 1 hour is suggested. Recommended Dilutions: ELISA: 1/75,000-1/300,000. Western Blot: 1/2,000-1/20,000. Immunohistochemistry: 1/500. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
HSPC193 antibody, HsT17436 antibody, JV15-2 antibody, LDS1C antibody, LDS3 antibody, MADH3 antibody, madh3 antibody, madh3a antibody, smad3 antibody, wu:fa99e03 antibody, XSmad3 antibody, XenMLP antibody, Xmad3 antibody, madh3-A antibody, Madh3 antibody, AU022421 antibody, Smad 3 antibody, mad3 antibody, madh3b antibody, zgc:92234 antibody, SMAD family member 3 antibody, SMAD family member 3a antibody, SMAD family member 3 L homeolog antibody, SMAD family member 3b antibody, SMAD3 antibody, smad3a antibody, smad3.L antibody, Smad3 antibody, smad3b antibody
Background
Smad3 (also known as Mothers against decapentaplegic homolog 3 Mothers against DPP homolog 3, Mad3, hMAD-3, JV15-2 or hSMAD3) is a transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinase. These activators exert diverse effects on a wide array of cellular processes. The Smad proteins mediate much of the signaling responses induced by the TGF-beta superfamily. Briefly, activated type I receptor phosphorylates receptor-activated Smads (R-Smads) at their c-terminal two extreme serines in the SSXS motif, e.g. Smad2 and Smad3 proteins in the TGF-beta pathway, or Smad1, Smad5 or Smad8 in the BMP pathway. Then the phosphorylated RSmad translocated into nucleus, where they regulate transcription of target genes. Based on microarray and animal model experiments, Smad3 accounts for at least 80% of all TGF-beta- mediated response. Alternate names: JV15-2, MAD homolog 3, MAD-3, MADH3, Mad3, Mothers against DPP homolog 3, Mothers against decapentaplegic homolog 3, SMAD 3, SMAD family member 3, SMAD-3