Revision 5

#7244

Store at +4C

PathScan® Total SMAD2 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)

orders@cellsignal.com

Support:

877-678-TECH (8324)

Web:

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 Rabbit mAb Coated Microwells7153796 tests+4C
SMAD2/3 Mouse Detection mAb131541 eaGreen (Lyophilized)+4C
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated)133041 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.

The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

CST's PathScan® Total SMAD2 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of total SMAD2 protein. A SMAD2 Rabbit mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-SMAD2 proteins are captured by the coated antibody. Following extensive washing, SMAD2/3 Mouse mAb is added to detect both the captured phospho- and nonphospho-SMAD2 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of optical density for this developed color is proportional to the quantity of total SMAD2 protein.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

CST's PathScan® Total SMAD2 Sandwich ELISA Kit #7244 detects endogenous levels of total SMAD2 protein. As shown in Figure 1, both phospho- and nonphospho-SMAD2 proteins from untreated and TGFβ treated MV1LU cell lysates are detected by this kit. In Figure 3, Western blot analysis of protein captured in the SMAD2 Rabbit mAb #3109 coated microwell shows a single band corresponding to the SMAD2 protein. SMAD2 in Hela cells also can be detected by this kit (data not shown). 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).

Background References

Trademarks and Patents

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

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

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) • orders@cellsignal.com • Support: 877-678-TECH (8324) • info@cellsignal.com • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 5

#7244

PathScan® Total SMAD2 Sandwich ELISA Kit

Cell Signaling Technology Logo
Figure 1: Nonphospho and phospho-SMAD2 proteins from untreated and TGF-β treated Mv1Lu cells detected by PathScan® Total SMAD2 Sandwich ELISA kit #7244 showing similar optical density readings. OD 450 readings are shown in the top figure, while the corresponding Western blots using SMAD2 Mouse mAb #3103 (left panel) or Phospho-SMAD2 (Ser465/467) Antibody #3101 (right panel), are shown in the bottom figure.
ELISA Image 1: PathScan<sup>®</sup> Total SMAD2 Sandwich ELISA Kit
Figure 2: The relationship between protein concentration of lysates from untreated and TGF-β-treated Mv1Lu cells and kit assay optical density readings. After starvation, Mv1Lu cells (85% confluence) were treated with TGF-β (100 ng/ml) for 15-30 min at 37°C, and then lysed.
ELISA Image 2: PathScan<sup>®</sup> Total SMAD2 Sandwich ELISA Kit
Figure 3: Kit specificity demonstrated by Western blot analysis of the ELISA well captured protein is shown. Lysates were prepared from mink Mv1Lu cells and incubated in wells coated with capture antibody #3109. Wells were then washed, and captured protein was solubilized in SDS gel loading buffer. Mv1Lu lysate (lane 1) and captured protein (lane 2) were analyzed by Western blot using SMAD2 Mouse mAb #3119. A single band corresponding to the SMAD2 protein is detected in the captured material (lane 2).
ELISA Image 3: PathScan<sup>®</sup> Total SMAD2 Sandwich ELISA Kit
Orders: 877-616-CELL (2355) • orders@cellsignal.com • Support: 877-678-TECH (8324) • info@cellsignal.com • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.