Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

Phospho-HER2/ErbB2 Antibody Sampler Kit #9923

Kit Includes Quantity Applications Reactivity MW (kDa) Source
Phospho-HER2/ErbB2 (Tyr877) Antibody # 2241 40 microliters W IHC-P H (M) (R) 185 Rabbit
Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb # 2243 40 microliters W IHC-P IHC-F H 185 Rabbit
HER2/ErbB2 Antibody # 2242 40 microliters W IHC-P H 185 Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074 100 microliters Goat

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)
Reactivity Key:  H=Human  M=Mouse  R=Rat

Specificity / Sensitivity

All antibodies in this kit detect endogenous levels of their targeted tyrosine-phosphorylated ErbB2 proteins. Each phospho-HER2/ErbB2 antibody recognizes only the phosphorylated form of HER2/ErbB2 at the indicated sites. The control HER2/ErbB2 receptor antibody recognizes both phosphorylated and nonphosphorylated forms of HER2/ErbB2.

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-treated (100 ng/ml) for various times, using Phospho-HER2/ErbB2 (Tyr877) Antibody #2241 (upper) or HER2/ErbB2 Antibody #2242 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using HER2/ErbB2 Antibody #2242.

Western Blotting

Western Blotting

Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb #2243 (upper) or Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower). Phospho-HER2/ErbB2 (Tyr1221/1222) (6B12) Rabbit mAb specifically detects phosphorylated HER2/ErbB2 but not other phosphorylated tyrosine kinases.


Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells expressing human ErbB2, untreated or EGF-treated, using Phospho-HER2/ErbB2 (Tyr877) Antibody #2241 (A), Phospho-HER2/ErbB2 (Tyr1248) Antibody #2244 (B) and HER2/ErbB2 Antibody #2242 (C).

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-peptides (KLH-coupled) corresponding to residues surrounding Tyr877, Tyr1112 or Tyr1248 of human HER2/ErbB2. HER2/ErbB2 Antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues surrounding Tyr1222 of human HER2/ErbB2. Antibodies are purified by protein A and peptide affinity chromatography.

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 with the regulation of ErbB2 degradation by the c-Cbl-regulated proteolytic pathway as 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 areTyr1248 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, 6845-6857.
  2. Qian, X. et al. (1994) Proc. Natl. Acad. Sci. USA 91, 1500-1504.
  3. Dittadi, R. and Gion, M. (2000) J. Natl. Cancer Inst. 92, 1443-1444.
  4. Klapper, L.N. et al. (2000) Cancer Res. 60, 3384-3388.
  5. Kwon, Y.K. et al. (1997) J. Neurosci. 17, 8293-8299.

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