Revision 6

#7148Store at +4C

1 Kit

(96 assays)

Species Cross Reactivity

H

UniProt ID:

#P04626

Entrez-Gene Id:

#2064

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Color Storage Temp
HER2/ErbB2 Mouse mAb Coated Microwells 26047 96 tests +4C
Phospho-HER2/ErbB2 (Tyr1221/1222) Rabbit Detection mAb 14099 1 ea Green (Lyophilized) +4C
Anti-rabbit IgG, HRP-linked Antibody (ELISA Formulated) 13272 1 ea Red (Lyophilized) +4C
Detection Antibody Diluent 13339 11 ml Green +4C
HRP Diluent 13515 11 ml Red +4C
TMB Substrate 7004 11 ml +4C
STOP Solution 7002 11 ml +4C
Sealing Tape 54503 2 ea +4C
ELISA Wash Buffer (20X) 9801 25 ml +4C
ELISA Sample Diluent 11083 25 ml Blue +4C
Cell Lysis Buffer (10X) 9803 15 ml -20C

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

Description

CST's PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-HER2/ErbB2 (Tyr1221/1222) protein. A HER2/ErbB2 Mouse mAb has been coated onto the microwells. After incubation with cell lysates, both phospho- and nonphospho-HER2/Erb2 Receptor proteins are captured by the coated antibody. Following extensive washing, Phospho-HER2/ErbB2 (Tyr1221/1222) Antibody is added to detect the captured phospho-HER2/ErbB2 protein. 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 optical density for this developed color is proportional to the quantity of phospho-HER2/ErbB2 (Tyr1221/1222) protein.

*Antibodies in kit are custom formulations specific to kit.

Specificity/Sensitivity

CST's PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit detects endogenous levels of phospho-HER2/ErbB2 (Tyr1221/1222) protein. As shown in Figure 1, using PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit, a high level of phospho-HER2/ErbB2 (Tyr1221/1222) protein is detected in Calu-3 cells, and the levels are significantly reduced in Calu-3 cells treated with λ protein phosphastase. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

The ErbB2 (HER2) proto-oncogene encodes a 185 kDa transmembrane, receptor-like glycoprotein with intrinsic tyrosine kinase activity (1). While ErbB2 lacks an identified ligand, ErbB2 kinase activity can be activated in the absence of a ligand when overexpressed and through heteromeric associations with other ErbB family members (2). Amplification of the ErbB2 gene and overexpression of its product are detected in almost 40% of human breast cancers (3). Binding of the c-Cbl ubiquitin ligase to ErbB2 at Tyr1112 leads to ErbB2 poly-ubiquitination and enhances degradation of this kinase (4). ErbB2 is a key therapeutic target in the treatment of breast cancer and other carcinomas and targeting the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway is one potential therapeutic strategy. Phosphorylation of the kinase domain residue Tyr877 of ErbB2 (homologous to Tyr416 of pp60c-Src) may be involved in regulating ErbB2 biological activity. The major autophosphorylation sites in ErbB2 are Tyr1248 and Tyr1221/1222; phosphorylation of these sites couples ErbB2 to the Ras-Raf-MAP kinase signal transduction pathway (1,5).

  1. Muthuswamy, S.K. et al. (1999) Mol Cell Biol 19, 6845-57.
  2. Qian, X. et al. (1994) Proc Natl Acad Sci USA 91, 1500-4.
  3. Dittadi, R. and Gion, M. (2000) J Natl Cancer Inst 92, 1443-4.
  4. Klapper, L.N. et al. (2000) Cancer Res 60, 3384-8.
  5. Kwon, Y.K. et al. (1997) J Neurosci 17, 8293-9.

Background References

    Cross-Reactivity Key

    H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

    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.

    Revision 6
    #7148

    PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit

    PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit: Image 1 Expand Image
    Figure1: Phosphorylation of HER2/ErbB2 at Tyr1221/1222 detected by PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA kit #7148. HER2/ErbB2 receptor kinases in Calu-3 cells are known to be constitutively activated. Treatment of Calu-3 cell lysates with λ protein phosphatase (37ºC for 60 minutes) abolished the phosphorylation of HER2/ErbB2 in the lysates shown in both sandwich ELISA and Western analysis. OD 450 readings are shown in the top figure, while the corresponding Western blots, using Phospho-HER2/ErbB2 (Tyr1221/1222) Antibody #2243 (right panel) or HER2/ErbB2 Antibody #2242 (left panel), is shown in the bottom figure.
    PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit: Image 2 Expand Image
    Figure 2: The relationship between protein concentration of untreated and LPP-treated Calu-3 cell lysates (see the legend of Figure 1) and kit assay optical density readings.