Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Tyrosine Kinase / Adaptors

Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) #8564

Applications Reactivity Sensitivity Isotype
IF-IC H M R Endogenous Rabbit

Applications Key:  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) detects endogenous levels of Met only when phosphorylated at Tyr1234/1235. This antibody may cross-react with overexpressed tyrosine phosphorylated Src proteins in western blot.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr1234/1235 of human Met protein.

IF-IC

IF-IC

Confocal immunofluorescent analysis of MKN-45 cells, untreated (left) or treated with Su11274 (right), using Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb (Alexa Fluor® 594 Conjugate) (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 594 fluorescent dye and tested in-house for direct immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Met (Tyr1234/1235) (D26) XP® Rabbit mAb #3077.

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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