Cell Signaling Technology

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-Met (Tyr1234/1235) Blocking Peptide #1645

Description

This peptide is used to block Phospho-Met (Tyr1234/1235) (D26) Rabbit mAb #3077 reactivity in western and immunohistochemistry protocols.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-Met (Tyr1234/1235) (D26) Rabbit mAb #3077 signal in western blotting and immunohistochemistry.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded HCC827 xenograft using Phospho-Met (Tyr1234/1235) (D26) Rabbit mAb #3077 in the presence of control peptide (left) or Phospho-Met (Tyr1234/1235) Blocking Peptide (right).

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in western and immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the relevant product data sheet.

Background

Met, a high affinity tyrosine kinase receptor for hepatocyte growth factor (HGF, also known as scatter factor) is a disulfide-linked heterodimer made of 45 kDa α- and 145 kDa β-subunits (1,2). The α-subunit and the amino-terminal region of the β-subunit form the extracellular domain. The remainder of the β-chain spans the plasma membrane and contains a cytoplasmic region with tyrosine kinase activity. Interaction of Met with HGF results in autophosphorylation at multiple tyrosines, which recruit several downstream signaling components, including Gab1, c-Cbl, and PI3 kinase (3). These fundamental events are important for all of the biological functions involving Met kinase activity. The addition of a phosphate at cytoplasmic Tyr1003 is essential for Met protein ubiquitination and degradation (4). Phosphorylation at Tyr1234/1235 in the Met kinase domain is critical for kinase activation. Phosphorylation at Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Research studies have shown that altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon, and breast. Thus, investigators have concluded that Met is an attractive potential cancer therapeutic and diagnostic target (6,7).

  1. Cooper, C.S. et al. (1984) Nature 311, 29-33.
  2. Bottaro, D.P. et al. (1991) Science 251, 802-4.
  3. Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
  4. Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
  5. Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
  6. Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
  7. Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.

Application References

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Companion Products

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

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