Anti-Phospholipid Screen ELISA Kit
Anti-Phospholipid Screen ELISA Kit Developed and Manufactured by Medipan
Size: 1×96 wells
Sensitivity: 1 U/ml
Dynamic Range: 1 – 300 U/ml
Incubation Time: 2.5 hours
Sample Type: Serum
Sample Size: 10 µL
Alternative Names: Human Anti-Phospholipid Screen ELISA, Human Phospholipid Antibody Screen ELISA
For Research Use Only
Assay Principle
The Anti-Phospholipid Screen ELISA Assay Kit is an enzyme immunoassay for the quantitative determination of IgG and / or IgM antibodies to negatively charged phospholipids.
The antibodies of the standards, controls and diluted patient samples react with an antigen complex consisting of cardiolipin, phosphatidyl-serine, -inositol, -ethanolamine, phosphatidic acid and the cofactor beta-2 GP-I immobilized on the solid phase of microtiter plates. The use of highly purified human beta-2 GP-I guarantees the specific binding of autoimmune related phospholipid antibodies of the specimen under investigation. Following an incubation period of 60 min at room temperature, unbound sample components are removed by a wash step.
The bound antibodies react specifically with anti-human-IgG or anti-human-IgM conjugated to horseradish peroxidase (HRP) within the incubation period of 30 min at room temperature (RT). Excessive conjugate is separated from the solid-phase immune complexes by the following wash step.
HRP converts the colorless substrate solution of 3,3’,5,5’-tetramethyl¬benzidine (TMB) added into a blue product. The enzyme reaction is stopped by dispensing an acidic solution into the wells after 15 min at room temperature turning the solution from blue to yellow.
The optical density (OD) of the solution at 450 nm is directly proportional to the amount of specific antibodies bound. The standard curve is established by plotting the antibody concentrations of the calibrators (x-axis) and their corresponding OD values (y-axis) measured. The concentration of antibodies of the specimen is directly read off the standard curve. Evaluating the test by a semi-quantitative method using a cut-off calibrator is also possible.
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