Product Pathways - PathScan ELISA
PathScan® Phospho-Met (panTyr) Sandwich ELISA Antibody Pair #7334
| Kit Includes | Volume | Cap Color |
|---|---|---|
| Met Rabbit Capture Antibody (100X) | 0.4 milliliters | Pink |
| Phospho-Tyrosine Mouse Detection Antibody (100X) | 0.4 milliliters | Blue |
| Anti-Mouse IgG HRP-Linked Antibody (1000X) | 0.04 milliliters | Yellow |
Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.
Species Cross-Reactivity
H
Reactivity Key: H=Human
Description
CST's PathScan® Phospho-Met (panTyr) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Met (panTyr) Sandwich ELISA Kit #7333. 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 Met Rabbit Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by Phospho-Tyrosine Mouse Detection Antibody and HRP-conjugated Secondary 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-Met (panTyr) protein.*Antibodies in this kit are custom formulations specific to the kit.
Sandwich ELISA
The relationship between protein concentration of phospho and nonphospho lysates and the absorbance at 450 nm is shown. Unstarved HCC827 cells were cultured (85% confluence) and lysed with or without addition of phosphatase inhibitor to the lysis buffer (phospho or nonphospho lysate).
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. Addition of a phosphate at cytoplasmic Tyr1003 is essential for ubiquitination and Met protein degradation (4). Phosphorylation of Tyr1234/1235 in the Met kinase domain is critical to kinase activation. Phosphorylation of Tyr1349 in the Met cytoplasmic domain provides a direct binding site for Gab1 (5). Altered Met levels and/or tyrosine kinase activities are found in several types of tumors, including renal, colon and breast. Thus, Met is an attractive cancer therapeutic and diagnostic target (6).
- Weidner, K.M. et al. (1993) J. Cell Biol. 121, 145-154.
- Park, M. et al. (1986) Cell 45, 895-904.
- Bardelli, A. et al. (1997) Oncogene 15, 3103-3111.
- Taher, T.E. et al. (2002) J. Immunol. 169, 3793-3800.
- Schaeper, U. et al. (2000) J. Cell Biol. 149, 1419-1432.
- Traxler, P. et al. (2001) Med. Res. Rev. 21, 499-512.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
