ANA Screen ELISA Kit
ANA Screen ELISA Kit is for research use only.
Size: 1 × 96 wells
Incubation Time: 1 hour 15 minutes
Sample Type: Serum, Plasma
Sample Size: 10 µL
Alternative Names: Antinuclear Antibody Screen ELISA, Human ANA Screen ELISA
Controls Included
Assay Background
The anti-nuclear antigen autoantibodies represent a large family of non-organ- and non-species-specific autoantibodies, detection of which is of great importance and widely used in laboratory diagnosis of systemic rheumatic autoimmune diseases. From the laboratory point of view, systemic autoimmune diseases are characteri4ed by the presence of anti-nuclear antibodies (ANAs). ANA is the first autoantibody test ordered for patients with suspected systemic autoimmune disorders. Despite its considerable use in diagnosis of these autoimmune disorders, ANA is often detected in healthy subjects without clinical presentation of any autoimmune disorder. It has recently been suggested that detection of ANA in such healthy subjects is a risk factor for development of connective tissue disease.
ANAs can be assessed by a variety of methods with the immunofluorescence assay (IFA) being considered the gold standard method. IFA positivity for ANA indicates the presence of autoantibodies directed against various nuclear antigens (DNA, histones, non-histonic proteins, nuclear antigens, etc.) or cytoplasmic antigens. Significantly high-titer positivity for ANAs should be further investigated via testing for anti-ENA, anti-dsDNA and anti-CENP-B autoantibodies. Positivity for ANAs and for one or more specific tests for anti-ENA, anti-dsDNA, and/or anti-CENP is highly suggestive of systemic autoimmune disorders: systemic lupus erythematosus (SLE), Sjogren’s Syndrome (SS), progressive systemic sclerosis (PSS), dermatomyositis/polymyositis (DM/PM), and/or mixed connective tissue disease (MCTD).
The most useful and most commonly tested anti-ANA autoantibodies are anti- Sm (Smith), anti-RNP/Sm, anti-Scl70, anti SS-A/Ro, anti SS-B/La, anti-Jo1, anti-CENP-B, histones and anti-dsDNA.
Assay Principle
The ANA Screen test is an indirect enzyme immunometric assay (ELISA) based on the binding of present antibodies with Sm, RNP/Sm, SS-A (Ro), SS-B (La), Scl-70, Jo1, U1-SmRNP, CENP-B, dsDNA and Histones antigens coated on the microplates. Any antibodies present in calibrators, controls or prediluted patient samples bind to the inner surface of the wells. After a 30 minutes incubation the microplate is washed with wash buffer for removing non-reactive serum components. An anti-human-IgG horseradish peroxidase conjugate solution recognizes IgG class antibodies bound to the immobilized antigens. After a 30 minutes incubation any excess enzyme conjugate, which is not specifically bound is washed away with wash buffer. A chromogenic substrate solution containing TMB is dispensed into the wells. After 15 minutes of incubation the color development is stopped by adding the stop solution. The solution turns yellow at this point. The level of color is directly proportional to the concentration of IgG antibodies present in the original sample.
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