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12052
SMAD2/3 Control Cell Extracts
Experimental Controls
Cell Extract Kit

SMAD2/3 Control Cell Extracts #12052

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  1. WB
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).

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).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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To Purchase # 12052
Cat. # Size Qty. Price
12052S
100 µl  (10 western blots)