Rat Anti Nuclear Antigens (ANA/ENA) IgG specific ELISA Kit

cat#650-230-ANA

INTENDED USE

The Rat ANA (Anti-Nuclear Antigens) IgG ELISA Kit is animmunoassay suitable for quantifying or titering IgG antibodyactivity specific for extractable nuclear antigens (ENA) in serumor plasma. Other biological fluids, including tissue culturemedium, may be validated for use. For research use only(RUO), not for diagnosis, cure or prevention of the disease.

GENERAL INFORMATION

Antibodiesreactive withautologousnuclearcomponents,such as DNAand histones,can representanautoimmunebasis forpathologicalconditions such as systemic lupus erythematosis (SLE) inhumans, and in mice homozygous for the lymphoproliferationspontaneous mutation (Faslpr), a systemic autoimmunity withmassive lymphadenopathy associated with proliferation ofaberrant T cells, arthritis and immune complexglomerulonephritis. These conditions include elevated levels ofanti-dsDNA and other anti-nuclear antigen (ANA) antibodieswhich often increase as the animal ages. Also, the expanded usein the drug industry of biological modifiers has been associatedwith production of autoantibodies, of which mice, and possiblyalso other hosts such as humans, monkeys and rats, aresusceptible. A prototype disease in mice is lupus caused by thedrug minocycline, with elevated anti-dsDNA among otherautoantibodies and pathological conditions.Recent investigations have focused on the role of innate immunemechanisms as underlying cause of the ANA type autoimmunity;these may be induced by drugs, including vaccines andadjuvants, with aging, or with other health conditions.

PRINCIPLE OF THE TEST

The Rat ANA IgG ELISA kit is based on the binding of rat ANAIgG in samples to ENA immobilized on the microwells, and ANAIgG antibody is detected by anti-Rat IgG-specific antibodyconjugated to HRP (horseradish peroxidase) enzyme. After awashing step, chromogenic substrate (TMB) is added and coloris developed by the enzymatic reaction of HRP on the substrate,which is directly proportional to the amount of ANA IgG presentin the sample. Stopping Solution is added to terminate thereaction, and absorbance at 450nm is then measured using anELISA microwell reader. The activity of rat IgG antibody insamples is calculated relative to ANA calibrators.

Specificity

Purified mammalian ENA is used to coat the microwells; thus theassay is specific for antibodies directed to ENA. The anti-Rat IgGHRP conjugate reacts specifically with rat IgG; IgA, IgM and IgEclass antibodies would not be measured above backgroundsignals.