A solid phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal antibody specific for c-Met (regardless of phosphorylation state) has been coated onto the wells of the multiwell plate strips provided. c-Met standard dilutions, control specimens, and unknown samples are incubated for 2 hours at RT. c-Met antigen binds to the immobilized (capture) antibody. After a wash, a anti-c-Met detection antibody specific for phosphorylated or non-phosphorylated protein is incubated for 1 hour at RT, which results in binding to the immobilized c-Met protein. An anti-rabbit IgG-HRP binds to the detection antibody to complete the four-member sandwich. The rection is visualized by TMB substrate. The intensity of the yellow color is directly proportional to the concentration of c-Met present in the original specimen. The optical density measured at 450 nm in the multiwell plate reader is used to calculate the concentration of c-Met. The unknown concentrations are calculated from the standard curve run with each assay.
Designed to quantify full length human c-Met protein in cell lysates. The assay does not detect soluble c-Met.
phospho-c-Met (pTyr1230/pTyr1234/pTyr1235) detects c-Met protein phosphorylated on tyrosines 1230, 1234, and 1235. The sensitivity is <0.25 units/mL, which is twice as sensitive as immunoblotting.
c-Met, a member of the tyrosine kinase superfamily, is the receptor for hepatocyte growth factor (also known as scatter factor). Many types of cancers exhibit sustained c-Met stimulation, overexpression, or mutation.
Features and Benefits
c-Met ELISA is a fast (total time 3.5 hours), sensitive (minimum detectable concentration >0.4 ng/ml) and specific alternative to immunoblotting and RIA. The ease of run is facilitated by precoated plates.