c-Met (Soluble) Human ELISA Kit

Catalog number:  KHO2031

 Related applications: Protein Assays and Analysis

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The Human c-Met (Soluble) ELISA research-use-only kit is designed to detect and quantify the level of soluble human c-Met protein in serum, plasma, and cell culture supernatants using 96-well plates and a microplate reader. The assay will recognize both natural and recombinant soluble human c-Met.

Performance characteristics
• Sensitivity: <0.5 ng/mL
• Standard curve range: 0.78–50 ng/mL
• Sample types: serum, plasma, cell culture supernatants
• Species cross-reactivity: human
• Sample volume: 100 μL (1:2 to 1:100)
• Total assay incubation time: 4 hrs

Principle of the method
The Human c-Met (Soluble) kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA). A highly purified antibody has been coated onto the wells of the microtiter strips provided. During the first incubation, standards of known soluble human c-Met content, controls, and unknown samples are pipetted into the wells. After washing, biotinylated secondary antibody is added. After washing, streptavidin-peroxidase (enzyme) is added. This binds to the biotinylated antibody to complete the four-member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of soluble human c-Met present in the original specimen.

Background information
c-Met, a member of the tyrosine kinase superfamily, is the receptor for hepatocyte growth factor, also known as scatter factor (HGF/SF). Cells expressing c-Met include epithelial cells, endothelial cells, blood cells of various types, and glomerular mesenchymal cells. Sources of HGF/SF include mesenchymal cells, mesanglial cells, endothelial cells, macrophages, and tumor cells.

HGF/SF binding to c-Met stimulates receptor dimerization and the phosphorylation of numerous residues within the receptor’s cytoplasmic domain. Phosphorylation of tyrosines 1234 and 1235 of c-Met is required for activation of the receptor’s tyrosine kinase activity. c-Met phosphorylation also generates docking sites for numerous signaling molecules and stimulates receptor internalization via clathrin-coated vesicles. Signaling proteins that are phosphorylated and/or localized in response to c-Met phosphorylation include: Grb2, Shc, Cbl, Crk, cortactin, paxillin, GAB1, PI-3 K, FAK, Src, Ras, ERK1 and 2, JNK, PLC-γ, AKT, and STAT3. HGF/SF stimulation of c-Met expressing cells enhances proliferation, migration, morphogenesis, and protease synthesis, characteristics that are associated with invasive cell phenotype.

Soluble c-Met is a truncated form of the c-Met membrane receptor. It can be cleaved by proteases and released from the lipid bilayer in a process known as ectodomain shedding. Many transmembrane proteins are released through this shedding process and it is a normal part of development which when defective can cause a number of pathologies. The soluble form of the c-Met receptor is smaller than the membrane bound receptor, contains the extracellular region of the receptor, and is able to bind the HGF ligand.

Detect low-expressing proteins and use less sample
ELISA kits designed to measure intracellular signaling targets are typically 2–10 times more sensitive than western blotting. The improved sensitivity enables you to detect low-expressing proteins that otherwise may not be distinguishable from background. In addition, the amount of sample needed to run the assay is less than what is needed for western blots.

Each ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See product insert for more information on validation.

Related links
Learn more about ELISA kits
Learn more about other immunoassays
For Research Use Only. Not for use in diagnostic procedures.


Protein Family: Kinases & Inhibitors
Assay Incubation Time: 4 hr
Assay Range: 0.78-50 ng/mL
Assay Sensitivity: < 0.5 ng/mL
Detection Method: Colorimetric
For Use With (Equipment): Microplate Reader
Former Identifiers: Novex
Gene Aliases: AUTS9, DFNB97, HGFR, RCCP2, c-Met
Gene ID: 4233
Gene Name: MET proto-oncogene, receptor tyrosine kinase
Gene Symbol: MET
Immunoassay Kit Format: Coated ELISA Kit
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 tests
Protein Name: Hepatocyte growth factor receptor
Protein Subtype: Receptor Tyrosine Kinases
Sample Type (Specific): Cell Culture Supernatants, Plasma, Serum
Sample Volume: 100 µL
Shipping Condition: Wet Ice
Target (Kit Named): c-Met
Target Species (Named): Human
Target Species (Validated): Human
Target Sub-Specificity: Soluble
UniProt ID: P08581

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.


Manuals & protocols