PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit #7348
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Supporting Data
| REACTIVITY | H M Mi |
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- Mi-Mink
Product Information
Product Description
The PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of SMAD2 when phosphorylated at Ser465/467. A SMAD2 Mouse monoclonal Antibody has been coated onto the microwells. After incubation with cell lysates, SMAD2 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a Phospho-SMAD2 (Ser465/467) Detection Antibody is added to detect serine phosphorylation of the captured SMAD2 protein. Anti-rabbit IgG, HRP-Linked Ab is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of SMAD2 phosphorylated at Ser465/467.
*Antibodies in this kit are custom formulations specific to kit.
Protocol
Specificity / Sensitivity
CST's PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit #7348 detects SMAD2 when phosphorylated at Ser465/467. As shown in Figure 1, a significant induction of SMAD2 phosphorylation at Ser465/467 can be detected in TGF-β-treated HaCaT cells using the PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit #7348. The level of total SMAD2 (phospho and nonphospho) remains unchanged as shown by western analysis and by PathScan® Total SMAD2 Sandwich ELISA Kit #7244 (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Species Reactivity:
Human, Mouse, Mink
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).
- 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.
- Wrana, J.L. (2000) Sci STKE 2000, re1.
- Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.
- Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.
Product Citations: 1
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