Cell Signaling Technology

Product Pathways - TGF-beta/Smad Signaling

Smad2/3 Control Cell Extracts #12052

Description

Nonphosphorylated Smad2/3 Control Cell Extracts: Total cell extracts from HT-1080 cells, serum-starved overnight without treatment, serve as a negative control. Supplied in SDS Sample Buffer. Phosphorylated Smad2/3 Control Cell Extracts: Total cell extracts from HT-1080 cells, serum-starved overnight and treated with hTGF-β3 #8425 (10 ng/ml, 30 min), serve as a positive control. Supplied in SDS Sample Buffer.

Western Blotting

Western Blotting

Western blot analysis of Smad2/3 Control Cell Extracts from HT-1080 cells, untreated (-) or treated with hTGF-β3 #8425 (10 ng/ml, 30 min; +), using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) or Smad2 (86F7) Rabbit mAb #3122 (lower).

Western Blotting

Western Blotting

Western blot analysis of Smad2/3 Control Cell Extracts from HT-1080 cells, untreated (-) or treated with hTGF-β3 #8425 (10 ng/ml, 30 min; +), using Phospho-Smad3 (Ser423/425) (C25A9) Rabbit mAb #9520 (upper) or Smad3 (C67H9) Rabbit mAb #9523 (lower).

Applications

As controls, we recommend using 10 μl of nonphosphorylated and phosphorylated Smad2/3 Control Cell Extracts. Boil for 2 minutes in the original tube, then load 10 μl per lane.

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).

  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. (1998) Genes Dev. 12, 2445-2462.
  6. Wu, G. et al. (2000) Science 287, 92-97.
  7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-1647.
  8. Moustakas, A. et al. (2001) J. Cell Sci. 114, 4359-4369.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products