Product Pathways - TGF-beta/Smad Signaling
Smad2 (D43B4) XP® Rabbit mAb #5339
|W IP IF-IC F ChIP||H M R Mk||Endogenous||60||Rabbit IgG|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Smad2 (D43B4) XP® Rabbit mAb detects endogenous levels of total Smad2 protein. This antibody does not cross-react with Smad3.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of mouse Smad2 protein.
Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb.
Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or treated with hTGF-β3 #8425 (right), using Smad2 (D43B4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either 10 μl of Smad2 (D43B4) XP® Rabbit mAb #5339 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 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).
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- Attisano, L. and Wrana, J.L. (1998) Curr. Opin. Cell Biol. 10, 188-194.
- Derynck, R. et al. (1998) Cell 95, 737-740.
- Massague, J. (1998) Annu. Rev. Biochem. 67, 753-791.
- Whitman, M. (1998) Genes Dev. 12, 2445-2462.
- Wu, G. et al. (2000) Science 287, 92-97.
- Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-1647.
- Moustakas, A. et al. (2001) J. Cell Sci. 114, 4359-4369.
- Katsu, K. et al. (2013) Dev Biol 374, 71-84. Applications: Western Blotting
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For Research Use Only. Not For Use In Diagnostic Procedures.