Revision 11

#7348

Store at +4C

PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit

1 Kit

(96 assays)

Species Cross Reactivity:
H M Mi

UniProt ID:
#Q15796

Entrez-Gene Id:
#4087

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Color Storage Temp
SMAD2 Mouse mAb Coated Microwells5890896 tests+4C
Phospho-SMAD2 (Ser465/467) Rabbit Detection mAb137671 eaGreen (Lyophilized)+4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated)132721 eaRed (Lyophilized)+4C
Detection Antibody Diluent1333911 mlGreen+4C
HRP Diluent1351511 mlRed+4C
TMB Substrate700411 ml+4C
STOP Solution700211 ml+4C
Sealing Tape545032 ea+4C
ELISA Wash Buffer (20X)980125 ml+4C
ELISA Sample Diluent1108325 mlBlue+4C
Cell Lysis Buffer (10X)980315 ml-20C

Kit contents scale proportionally with size, except sealing tape.
Example: The V1 kit contains 5X the listed quantities above, but will exclude the sealing tape.

For the “C” and “V” kits, the supplied 96-well strip plate consists of twelve 8-well strips in a support frame. This enables custom plate configurations.

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.

Specificity/Sensitivity

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 this Sandwich ELISA Kit #7348. The level of total Smad2 (phospho and nonphospho) remains unchanged as shown by western analysis (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.

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).

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 11

Cell Signaling Technology Logo
Figure 1. Treatment of HaCaT cells with TGF-β stimulates phosphorylation of SMAD2 at Ser465/467, detected by PathScan® Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit #7348, but does not affect the level of total SMAD2 protein detected by a Total SMAD2 Sandwich ELISA. The absorbance readings at 450 nm are shown in the top figure, while the corresponding western blots using SMAD2 (86F7) Rabbit mAb #3122 (left panel) or Phospho-SMAD2 (Ser465/467) (138D4) Rabbit mAb #3108 (right panel) are shown in the bottom figure.
ELISA Image 1: PathScan<sup>®</sup> Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit
Figure 2. The relationship between the protein concentration of lysates from untreated and TGF-β-treated HaCaT cells and the absorbance at 450 nm is shown. Cells (85% confluence) were treated with 10 ng/ml TGF-β for 30 min. at 37oC.
ELISA Image 2: PathScan<sup>®</sup> Phospho-SMAD2 (Ser465/467) Sandwich ELISA Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.