Description
Intended Uses: This Sphingomyelin IgG/M ELISA is a solid phase enzyme immunoassay with highly purified sphingomyelin and native human beta2-glycoprotein I for the quantitative and qualitative detection of IgG and/or IgM antibodies against sphingomyelin in human serum. These antibodies recognize specific epitopes on a complex composed out of sphingomyelin and beta2-glycoprotein I.
Principle of the Assay: Serum samples diluted 1:101 are incubated in the microplates coated with the specific antigen. 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 the sample.
Background: Antibodies against sphingomyelin belong to the group of anti-phospholipid antibodies such as cardiolipin, phosphatidyl- serine, -inositol, -choline and phosphatidic acid. Phospholipids are components of biological membranes