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7896
PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit
ELISA Kits
ELISA Kit

PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit #7896

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PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit: Image 1

Figure 1: Constitutive phosphorylation of Met in HCC827 cells lysed in the presence of phosphatase inhibitors (phospho lysate) is detected by PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit #7896 (top, right). In contrast, a low level of phospho-Met protein is detected in HCC827 cells lysed without addition of phosphatase inhibitors* (nonphospho lysate). Similar levels of total Met protein from either nonphospho or phospho lysates are detected by PathScan® Total Met Sandwich ELISA Kit #7242 (top, left). Absorbance at 450 nm is shown in the top figure, while the corresponding Western blots using Phospho-Met (Tyr1349) Rabbit mAb #3133 (right) or a total Met Antibody #4560 (left) are shown in the bottom figure. *Phosphatase inhibitors include sodium pyrophosphate, β-glycerophosphate and Na3VO4.

PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit: Image 2

Figure 2: The relationship between protein concentration of phospho or 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 (phospho or nonphospho lysate).

Inquiry Info.# 7896

Important Ordering Details

Custom Ordering Details: When ordering five or more kits, please contact us for processing time and pricing at [email protected].

Supporting Data

REACTIVITY

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Description

The PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of Met when phosphorylated at Tyr1349. A Met rabbit antibody has been coated onto the microwells. After incubation with cell lysates, phospho-Met (Tyr1349) is captured by the coated antibody. Following extensive washing, a biotinylated phospho-Met (Tyr1349) rabbit detection antibody is added to detect the captured phospho-Met protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The absorbance at 450nm is proportional to the quantity of Met phosphorylated at Tyr1349.

Antibodies in kit are custom formulations specific to kit.

MW (kDa) 145

Specificity / Sensitivity

CST's PathScan® Phospho-Met (Tyr1349) Sandwich ELISA Kit detects endogeneous levels of Met when phosphorylated at Tyr1349 (see Figure 1). The kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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PathScan is a trademark of Cell Signaling Technology, Inc.