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Product Pathways - TGF-beta/Smad Signaling

Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108

Item# Description List Price Web Price Qty
3108L Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb - 300 µl $1,028.00
$925.20
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3108S Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb - 100 µl $439.00
$395.10
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3108T Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb - 20 µl $174.00
$156.60
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Mink Endogenous 60 Rabbit IgG

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting

Protocols

Specificity / Sensitivity

Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb detects endogenous levels of Smad2 only when dually phosphorylated at serines 465 and 467, and may detect Smad3 phosphorylated at the equivalent sites. This antibody does not cross-react with other Smad-related proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser465/467 of human Smad2.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or TGF-beta treated HeLa and NIH/3T3 cells, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (upper), or Smad2 Antibody #3102 (lower).

Background

Members of the Smad family of signal transduction molecules are components of a critical intracellular pathway that transmit 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-β, Smad2 and Smad3 become phosphorylated at their carboxy-termini (Ser465/467 on Smad2; Ser423/425 on Smad3) by the receptor kinase TGF-β R1 (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, 465-71.
  2. Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.
  3. Derynck, R. et al. (1998) Cell 95, 737-40.
  4. Massagué, J. (1998) Annu Rev Biochem 67, 753-91.
  5. Whitman, M. (1998) Genes Dev 12, 2445-62.
  6. Wu, G. et al. (2000) Science 287, 92-7.
  7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
  8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.
  9. Abdollah, S. et al. (1997) J Biol Chem 272, 27678-85.
  10. Souchelnytskyi, S. et al. (1997) J Biol Chem 272, 28107-15.
  11. Liu, X. et al. (1997) Proc Natl Acad Sci U S A 94, 10669-74.

Application References

  • Kong, B. et al. (2010) Oncogene 29, 5146-58. Applications: Western Blotting.

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