Cell Signaling Technology

Product Pathways - Tyrosine Kinase/ Adaptors

HER2/ErbB2 (29D8) Rabbit mAb #2165

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IHC-F IF-IC F H Endogenous 185 Kda Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

HER2/ErbB2 (29D8) Rabbit mAb detects endogenous levels of total ErbB2 protein. This antibody does not cross-react with related kinases.

Source / Purification

Monoclonal antibodies are produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to residues surrounding tyrosines 1248 of human ErbB2 protein.

Western Blotting

Western Blotting

Western blot analysis of cell extracts from various cell lines, using HER2/ErbB2 (29D8) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using HER2/ErbB2 (29D8) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using HER2/ErbB2 (29D8) RmAb in the presence of control peptide (left) or HER2/ErbB2 Blocking Peptide #1059 (right).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded SKBr3 (high HER2) (left), MDA-MB-453 (moderate HER2) (middle) and MDA-MB-468 (low HER2) (right), using HER2/ErbB2 (29D8) Rabbit mAb.

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen SKOV-3 xenograft using HER2/ErbB2 (29D8) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of MCF7 cells using HER2/ErbB2 (29D8) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF-7 cells labeled with HER2/ErbB2 (29D8) Rabbit mAb (red). Blue= DRAQ5™ fluorescent DNA dye.

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.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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