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PhosphoSitePlus® Resource

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REACTIVITY
Product Includes Volume Solution Color
Smad2/3 Antibody Coated Microwells 96 tests
Phospho-Smad3 (Ser423/425) Rabbit Detection mAb 1 ea Green (Lyophilized)
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
TMB Substrate 7004 11 ml Colorless
STOP Solution 7002 11 ml Colorless
Sealing Tape 2 sheets
ELISA Wash Buffer (20X) 25 ml Colorless
ELISA Sample Diluent 25 ml Blue
Cell Lysis Buffer (10X) 9803 15 ml Yellowish

Product Description

The PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that recognizes endogenous levels of Smad3 (Ser423/425) protein. A Smad2/3 Mouse Antibody has been coated on the microwells. After incubation with cell lysates, Smad3 proteins (phospho and nonphospho) are captured by the coated antibody. Following extensive washing, a Phospho-Smad3 (Ser423/425) Rabbit Detection Antibody is added to detect captured phospho-Smad3 (Ser423/425) proteins. Anti-rabbit IgG, HRP-linked Antibody 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 phospho-Smad3 (Ser423/425) proteins.

Antibodies in kit are custom formulations specific to kit.


Specificity / Sensitivity

PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit recognizes endogenous levels of phospho-Smad3 (Ser423/425) protein in human cells, as shown in Figure 1. The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.


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).


1.  Heldin, C.H. et al. (1997) Nature 390, 465-71.

2.  Attisano, L. and Wrana, J.L. (1998) Curr Opin Cell Biol 10, 188-94.

3.  Massagué, J. (1998) Annu Rev Biochem 67, 753-91.

4.  Whitman, M. (1998) Genes Dev 12, 2445-62.

5.  Wu, G. et al. (2000) Science 287, 92-7.

6.  Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.

7.  Derynck, R. et al. (1998) Cell 95, 737-740.

8.  Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.


Entrez-Gene Id 4088
Swiss-Prot Acc. P84022


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a trademark of Cell Signaling Technology, Inc.

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PathScan® Phospho-Smad3 (Ser423/425) Sandwich ELISA Kit