Revision 2

#12052Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB

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Product Information

Product Usage Information

Boil for 3 minutes prior to use. Load 10 μl of phosphorylated and nonphosphorylated Smad2/3 Control Cell Extracts per lane.

Storage

Supplied in SDS Sample Buffer: 62.5 mM Tris- HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM DTT, 0.01% w/v bromophenol blue or phenol red.
Store at –20°C, or at –80°C for long-term storage.

Product Description

Nonphosphorylated Smad2/3 Control Cell Extracts: Total cell extracts from HT-1080 cells, serum-starved overnight to 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 10 ng/ml hTGF-β3 #8425 for 30 min to serve as a positive control. Supplied in SDS Sample Buffer.

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 9; 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-SMADs dissociate from the receptor and form a heteromeric complex with SMAD4, initiating translocation of the heteromeric SMAD complex to the nucleus. Once in the nucleus, SMADs recruit a variety of DNA binding proteins that function to regulate transcriptional activity (6-8).

  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. Wrana, J.L. (2000) Sci STKE 2000, re1.
  7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
  8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

WB: Western Blotting

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

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Revision 2
#12052

SMAD2/3 Control Cell Extracts

Western Blotting Image 1: SMAD2/3 Control Cell Extracts Expand Image
使用 Phospho-SMAD2 (Ser465/467) (138D4) Rabbit mAb #3108(上图)或 SMAD2 (86F7) Rabbit mAb #3122(下图),对未经处理 (-) 或经过 hTGF-β3 #8425(10 ng/ml,30 分钟;+)处理的 HT-1080 细胞的 SMAD2/3 对照细胞提取物进行蛋白质印迹法分析。