ANA Screen Reagent – Jaj International

The ANA screen ELISA Assay Kit is an enzyme immunoassay for the semi-quantitative determination of IgG antibodies to nuclear and cytoplasmic antigens.

Antibodies of the controls and diluted patient samples react with nuclear and cytoplasmic antigens immobilized on the solid phase of microtiter plates. Use of complete HeLa nuclei enriched with recombinant and native antigens guarantees the specific binding of autoimmune antibodies of the specimen under investigation. Following an incubation period of 60 min at room temperature (RT, 18…25°C), unbound sample components are removed by a wash step.

The bound IgG antibodies react specifically with anti-human-IgG conjugated to horseradish peroxidase (HRP). Within the incubation period of 30 min at 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. Patient ratios are calculated by dividing the respective OD of the specimen with the calculated cut-off OD.

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