The Eagle Biosciences 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 Antigens complex consisting of cardiolipin, phosphatidyl-serine, -inositol, -ethanolamine, phosphatidic acid and the cofactor b2 GP-I immobilized on the solid phase of microtiter plates. The use of highly purified human b2 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. Sample Types: serum,. Range: 1-300 U/mL. Sensitivity: 1U/mL. Time: 2.5 hours ? APAS is an autoimmune disorder comprising such clinical symptoms like arterial or venous thrombosis, thrombocytopenia and recurrent fetal loss. Primary APAS as well as systemic lupus erythematosus (SLE) are characterized by the appearance of autoantibodies to negatively charged phospholipids (1). Although significance and pathological relevance of phospholipid antibodies are not completely revealed yet, the detection of such autoantibodies is widely established and plays an important role in the diagnostics of systemic autoimmune diseases. Unlike phospholipid antibodies which appear in some infectious disease patients autoimmune patients exhibit phospholipid antibodies that seem to recognize phospholipids in association with plasma protein cofactors such as b2 glycoprotein-I (b2 GP-I) (apolipoprotein H) (2,3). b2 GP-I, a serum protein with a molecular weight of 50 kDa, affects platelet aggregation and coagulation.The positively charged fifth domain of b2 GP-I interacts with negatively charged phospholipids such as Cardiolipin. This interaction results in conformational changes of the protein and the creation of new epitopes apparently recognized by autoimmune phospholipid autoantibodies.
Size
1 x 96 well
Concentration
n/a
Applications
RUO
Other Names
PHS31-K01 ASSAY EIA ELISA IMMUNOASSAY KIT, Anti Phospholipid APAS