Product Pathways - TGF-beta/Smad Signaling
Smad4 Antibody #9515
|9515S||100 µl (10 western blots)||---||In Stock||---|
|9515P||40 µl (4 western blots)||---||In Stock||---|
|9515||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||70||Rabbit|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, ChIP=Chromatin IP
Specificity / Sensitivity
Smad4 Antibody detects endogenous levels of total Smad4 protein.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the residues surrounding Pro278 of human Smad4. Antibodies are purified by protein A and peptide affinity chromatography.
Western blot analysis of extracts from COS, NIH3T3, PC12, and SK-N-MC cells, using Smad4 Antibody.
Western blot analysis of extracts from NIH/3T3 cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Smad4 siRNA I (Mouse Specific) (+), using Smad4 Antibody #9515 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The Smad4 Antibody confirms silencing of Smad4 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #3706 (7ng/ml) for 1 h and either 20 μl of Smad4 Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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 recepter-regulated Smads (R-Smads), which include Smad1, 2, 3, 5, 8; the common-mediator Smad (co-Smad), Smad4; and the antagonistic or inhibitory Smads (I-Smads), Smad6 and 7 (1-5). Briefly, activated type I receptors associate with specific R-Smads and phosphorylate them on a conserved SSXS motif at the carboxy-terminus of the proteins. The phosphorylated R-Smad dissociates from the receptor and forms a heteromeric complex with the co-Smad, Smad4, and together the complex moves to the nucleus. Once in the nucleus, Smads can target a variety of DNA binding proteins to regulate transcriptional responses (6-8).
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- Moustakas, A. et al. (2001) J. Cell Sci. 114, 4359-4369.
- Upton, P.D. et al. (2009) J Biol Chem 284, 15794-804. Applications: Western Blotting.
- Van Themsche, C. et al. (2010) Mol Cancer 9, 216. Applications: Western Blotting.
- Ding, Y. et al. (2010) J Biol Chem 285, 37909-19. Applications: Western Blotting.
- Long, X. and Miano, J.M. (2011) J Biol Chem 286, 30119-29. Applications: Western Blotting.
- Wang, J. et al. (2013) J Biol Chem 288, 10418-26. Applications: Chromatin IP.
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