c-Met (soluble) ELISA Kit, Human

Catalog number: KHO2031

Novex™  Related applications: Protein Assays and Analysis

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Description

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.

Specifications

Gene Aliases: AUTS9, HGFR, RCCP2, c-Met
Protein Subtype: Receptor Tyrosine Kinases
Accession Number: NP_000236.2, NP_001120972.1, NM_000245.2, NM_001127500.1
Shipping Condition: Wet Ice
Marker: c-Met (soluble)
Gene ID: 4233
Species: Human
Gene Symbol: MET
Sensitivity: < 0.5 ng⁄ml
Target Gene: met proto-oncogene (hepatocyte growth factor receptor)
Label or Dye: HRP (Horseradish Peroxidase)
Product Size: 96 assays
Concentration: 0.78-50 ng⁄ml
Sample Volume: 100 µl
Protein Family: Kinases & Inhibitors
Incubation Time: 4 hrs
Detection Method: Colorimetric
Research Category: Signal Transduction
Sample Type (Specific): Plasma, Serum, Supernatants
For Use With (Equipment): Microplate Reader

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.

Documents

Manuals & protocols