Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad2 (Ser465/467) Antibody #3101

Applications Reactivity Sensitivity MW (kDa) Source
W H M R (C) (Dm) (X) (Z) Endogenous 60 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  C=Chicken  Dm=D. melanogaster  X=Xenopus  Z=Zebra Fish
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Smad2 (Ser465/467) Antibody detects endogenous levels of Smad2 only when dually phosphorylated at Ser465 and Ser467, and may detect phosphorylated Smad3 at its equivalent site. This 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 Ser465/467 of human Smad2. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HepG2 cells treated with TGFbeta for the indicated times, using Phospho-Smad2 (Ser465/467) Antibody (upper) or Smad2 antibody (lower).

Background

Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmits TGF-β 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-beta, Smad2 and Smad3 become phosphorylated at their carboxyl termini by the receptor kinase (serines 465 and 467 on Smad2; serines 423 and 425 on Smad3) by TbetaR-I (9-11). Following phosphorylation, Smad2 and Smad3 form a heteromeric complex with the co-smad family member Smad4. These complexes are translocated to the nucleus where they bind DNA and regulate gene transcription.

  1. Heldin, C.H. et al. (1997) Nature 390, 465-471.
  2. Attisano, L. and Wrana, J.L. (1998) Curr. Opin. Cell Biol. 10, 188-194.
  3. Derynck, R. et al. (1998) Cell 95, 737-740.
  4. Massague, J. (1998) Annu. Rev. Biochem. 67, 753-791.
  5. Whitman, M. et al. (1998) Genes Dev. 12, 2445-2462.
  6. Wrana, J. (2000) Science 23, 1-9.
  7. Attisano, L. and Wrana, J. (2002) Science 296, 1646-1647.
  8. Moustakas, A. et al. (2001) J. Cell Sci. 114, 4359-4369.
  9. Abdollah, S. et al. (1997) J. Biol. Chem. 272, 27678-27685.
  10. Souchelnytskyi, S. et al. (1997) J. Biol. Chem. 272, 28107-28115.
  11. Liu, X. et al. (1997) Proc. Natl. Acad. Sci. 94, 10669-10674.

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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.

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