Product Pathways - TGF-beta/Smad Signaling
Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody #9511
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IF-IC F | H M R Mi X | Endogenous | 60 | Rabbit |
Applications Key:
W=Western Blotting
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mi=Mink
X=Xenopus
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad8 (Ser426/428) Antibody detects endogenous levels of Smad1 only when dually phosphorylated at serine 463 and serine 465, as well as Smad5 and Smad8 only when phosphorylated at the equivalent sites. The antibody does not cross-react with other Smad-related proteins.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser463/465 of human Smad5. Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from untransfected Xenopus cells and BMPRI/Smad1-transfected 293 cells, untreated or BMP-4-treated, using Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad8 (Ser426/428) Antibody. (Xenopus cells provided by Dr. Malcolm Whitman, Harvard University, Massachusetts.)
Flow Cytometry
Flow cytometric analysis of HeLa cells, untreated (blue) or BMP-treated (green), using Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad8 (Ser426/428) Antibody compared with a nonspecific negative control antibody (red).
IF-IC
Confocal immunofluorescent images of HeLa cells, untreated (left) or BMP-2-treated (right), labeled with Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody (green) and Keratin 18 (DC10) Mouse mAb #4548 (blue). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).
Background
Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5).
- Hogan, B.L. et al. (1996) Genes Dev. 10, 1580-1594.
- Hoodless, P.A. et al. (1996) Cell 85, 489-500.
- Klemm, J.D. et al. (1998) Annu. Rev. Immunol. 16, 569-592.
- Kretzschmar, M. et al. (1997) Genes Dev. 11, 984-995.
- Whitman, M. (1998) Genes Dev. 12, 2445-2462.
Application References
- Kyung, A. et al. (2001) BMPR-1A signaling is required for the formation of the apical ectodermal ridge and dorsal-ventral patterning of the limb. Development 128, 4449-4461. This article references the use of Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody in the following applications: IHC-Paraffin
- Seto, H. et al. (2004) Distinct roles of Smad pathways and p38 pathways in cartilage-specific gene expression in synovial fibroblasts. J. Clin. Invest. 113, 718-726. This article references the use of Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody in the following applications: Western Blotting
- Jackson, L. F. et al. (2003) Defective valvulogenesis in HB-EGF and TACE-null mice is associated with aberrant BMP signaling. EMBO J. 22, 2704-2716. This article references the use of Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody in the following applications: IHC-P (paraffin)
- Watabe, T. et al. (2003) TGF-beta receptor kinase inhibitor enhances growth and integrity of embryonic stem cell-derived endothelial cells. J. Cell Biol. 163, 1303-1311. This article references the use of Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody in the following applications: Western Blotting
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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
