| Order number | Description | Quantity | Delivery time | CE |
|---|---|---|---|---|
| CO 10240 | Anti-Prothrombin IgM ELISA | 1 plate | 10-15 days |
Anti-Prothrombin IgM ELISAFor the detection and semi-quantitation of IgM antiprothrombin (aPT) antibodies in individuals with systemic lupus erythematosus (SLE) and lupus-like disorders (e.g., antiphospholipid syndrome). The test is performed as an indirect ELISA. Diluted serum or citrated plasma samples, calibrator, and controls are incubated in microwells coated with purified human prothrombin. Incubation allows the anti-prothrombin antibodies present in the samples to react with the immobilized antigen. After the removal of unbound proteins by washing, antibodies specific for human IgM labeled with horseradish peroxidase (HRP) are added forming complexes with the prothrombin bound antibodies. Following another washing step, the bound enzyme-antibody conjugate is assayed by the addition of a single solution containing tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) as the chromogenic substrate. Color develops in the wells at an intensity proportional to the concentration of anti-prothrombin antibodies. Results are obtained by reading the optical density (O.D. or absorbance) of each well in a spectrophotometer. Calibrator sera are provided with the IgM anti-prothrombin antibody concentration expressed in M units. The user has the option of running either a single point calibrator or a four-point calibration curve. For single point calibration, dividing the concentration value of the calibrator sera by the O.D. value of the calibrator provides a conversion factor. The O.D. values of the other samples are multiplied by the conversion factor to obtain IgM anti-prothrombin antibody concentrations in M units. For multipoint calibration, perform a linear regression analysis with calibrator values against calibrator O.D.'s. Controls and patient results are determined from the calibration curve. |
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