Product Pathways - PathScan ELISA
PathScan® Phospho-Met (panTyr) Sandwich ELISA Antibody Pair #7334
|Kit Includes||Volume||Cap Color|
|Met Capture Ab (100X)||0.4 ml||Pink|
|P-Tyrosine Detection Ab (100X)||0.4 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||0.04 ml||Yellow|
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Important Ordering Details
Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
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.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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).
- Cooper, C.S. et al. (1984) Nature 311, 29-33.
- Bottaro, D.P. et al. (1991) Science 251, 802-4.
- Bardelli, A. et al. (1997) Oncogene 15, 3103-11.
- Taher, T.E. et al. (2002) J Immunol 169, 3793-800.
- Schaeper, U. et al. (2000) J Cell Biol 149, 1419-32.
- Eder, J.P. et al. (2009) Clin Cancer Res 15, 2207-14.
- Sattler, M. and Salgia, R. (2009) Update Cancer Ther 3, 109-118.
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- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7242 PathScan® Total Met Sandwich ELISA Kit
- 7333 PathScan® Phospho-Met (panTyr) Sandwich ELISA Kit
For Research Use Only. Not For Use In Diagnostic Procedures.