ANA Screen ELISA Kit
ANA Screen ELISA Kit Developed and Manufactured Medipan
Size: 1×96 wells
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Antinuclear Antibody Screen ELISA, Human ANA Screen ELISA
For Research Use Only
Reference Values
Positive: ≥ 1.0
Negative: < 1.0
It is recommended that each laboratory establishes its own normal and pathological reference ranges, as usually done for other diagnostic parameters, too. Therefore, the above mentioned reference values provide a guide only to values which might be expected.
Specificity
Frequency distribution of ANA in ANAscreen: 97 unselected human sera from healthy donors were tested. All sera were found negative. This corresponds to a diagnostic specificity of 100%.
Assay Principle
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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