Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Antibody Pair #7817

Kit Includes Volume Cap Color
HER2/ErbB2 Capture Ab(100X) 0.4 ml Pink
HER2/ErbB2 (Y1221/ 1222) Detection Ab (100X) 0.4 ml Blue
Anti-rabbit IgG, HRP-linked Antibody 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Important Ordering Details

Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Protocols

Description

CST's PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Kit #7148. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The HER2/ErbB2 Capture Antibody is coated on a 96 well microplate in PBS overnight. After blocking, cell lysates are added followed by a Phospho-HER2/ErbB2 (Tyr1221/1222) Detection Antibody and anti-rabbit IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance 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

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

The relationship between protein concentration of untreated and λ phosphatase-treated Calu-3 cell lysates, and the absorbance at 450 nM using PathScan® Phospho-HER2/ErbB2 (Tyr1221/1222) Sandwich ELISA Antibody Pair #7817 is shown.

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.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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