Smad2 (D43B4) XP® Rabbit mAbProduct information
Smad2 (D43B4) XP® Rabbit mAb
Smad2 (D43B4) XP® Rabbit mAb
Product Pathways - TGF-beta/Smad Signaling
Smad2 (D43B4) XP® Rabbit mAb #5339
|5339S||100 µl||---||In Stock||---|
|5339T||20 µl||---||In Stock||---|
|5339||carrier free and custom formulation / quantity||email request|
|W||Human, Mouse, Rat, Monkey||Endogenous||60||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Directions For Use
For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
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.
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).
Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either Smad2 (D43B4) XP® Rabbit mAb #5339 or 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).
- Heldin, C.H. et al. (1997) Nature 390, 465-71.
- Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.
- Derynck, R. et al. (1998) Cell 95, 737-40.
- Massagué, J. (1998) Annu Rev Biochem 67, 753-91.
- Whitman, M. (1998) Genes Dev 12, 2445-62.
- Wu, G. et al. (2000) Science 287, 92-7.
- Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
- Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.
- Katsu, K. et al. (2013) Dev Biol 374, 71-84. Applications: Western Blotting.
- Tamiya, T. et al. (2013) J Immunol 191, 2360-71. Applications: Chromatin IP.
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.