Description
Intended Uses: This ELISA is a solid phase enzyme immunoassay for the separate qualitative detection of IgG and/or IgM antibodies against phospholipids in human serum. The assay employs highly purified human Prothrombin (Factor II), Thrombin, Cardiolipin, Phosphatidyl-choline, -ethanolamine, -inositol, -serine and Sphingomyelin.
Principle of the Assay: Serum samples diluted 1:101 are incubated in the microplates coated with the specific antigen. The antibodies, if present in the specimen, bind to the antigen. The unbound fraction is washed off in the following step. Afterwards anti-human immunoglobulins conjugated to horseradish peroxidase (conjugate) are incubated and react with the antigen-antibody complex of the samples in the microplates. Unbound conjugate is washed off in the following step. Addition of TMB-substrate generates an enzymatic colorimetric (blue) reaction, which is stopped by diluted acid (color changes to yellow). The intensity of color formation from the chromogen is a function of the amount of conjugate bound to the antigen-antibody complex and this is proportional to the initial concentration of the respective antibodies in sample.
Background: Antibodies against phospholipids, components of the biological membranes, are specific for phospholipids such as Cardiolipin, Phosphatidyl -inositol, -ethanolamine, -choline and Sphingomyelin