Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad2 (86F7) Rabbit mAb #3122

Applications Reactivity MW (kDa) Source Isotype
W IP IF-IC H Mk 60 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Smad2 (86F7) Rabbit mAb detects endogenous levels of total Smad2 protein. No cross reactivity was detected with Smad3.

Source / Purification

Rabbit monoclonal antibodies were prepared from spleens of rabbits immunized with a synthetic peptide (KLH-coupled) corresponding to residues surrounding Trp85 of human Smad2.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, H1975 and HT29 cell lines, using Smad2 (86F7) Rabbit mAb.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or TGF-β treated (right), using Smad2 (86F7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red)

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).

  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.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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