Monkey Anti dsDNA IgG ELISA Kit

cat#670-100-DNM  

INTENDED USE

The Monkey anti-dsDNA IgG ELISA Kit is an immunoassaysuitable for quantifying or titering IgG antibody activity specific fordouble-stranded DNA in serum or plasma. Other biological fluids,including tissue culture medium, may be validated for use. Forresearch use only, not for therapeutic use.

GENERAL INFORMATION

Antibodies reactive withautologous nuclear components,such as DNA and histones, canrepresent an autoimmune basisfor pathological conditions suchas systemic lupus erythematosis (SLE) in humans, and in micehomozygous for the lymphoproliferation spontaneous mutation(Faslpr), a systemic autoimmunity with massivelymphadenopathy associated with proliferation of aberrant Tcells, arthritis and immune complex glomerulonephritis. Theseconditions include elevated levels of anti-dsDNA and other antinuclear antibodies (ANA) which often increase as the animalages. Also, the expanded use in the drug industry of biologicalmodifiers has been associated with production of autoantibodies,of which mice, and possibly also other hosts such as humansand monkeys, are susceptible. A prototype disease in mice islupus caused by the drug minocycline, with elevated anti-dsDNAamong other autoantibodies and pathological conditions. Recentinvestigations have focused on the role of innate immunemechanisms, responding to the damage-associated molecularpatterns of dying cells, as underlying cause of the anti-dsDNAtype autoimmunity; these may be induced by drugs, includingvaccines and adjuvants, with aging, or with other conditions.

PRINCIPLE OF THE TEST

The Monkey anti-dsDNA IgG ELISA kit is based on the binding ofmonkey anti-dsDNA IgG in samples to dsDNA coated on theplate, and anti-dsDNA antibody is detected by anti-monkey IgGspecific antibody-HRP conjugate. After a washing step,chromogenic substrate (TMB) is added and color (blue) isdeveloped, which is directly proportional to the amount ofantibody present in the sample. Stopping Solution is added toterminate the reaction, and A450nm is then measured using anELISA reader. The activity and/or concentration of antibody insamples is calculated relative to supplied calibrators.

Specificity

Purified dsDNA is used to coat the microwells; thus the assay isspecific for antibodies directed to dsDNA. The anti-Monkey IgGHRP conjugate reacts specifically with monkey IgG subclassantibodies that bind to dsDNA on the plate. IgA, IgM, IgE andother IgG subclass antibodies would not be measured abovebackground signals.

Assay Sensitivity

The dsDNA coating level, HRP conjugate concentration and LowNSB Sample Diluent are optimized to differentiate anti-dsDNAIgG from background (non-antibody) signal with monkey serumsamples diluted 1:100.

Calibrator Values

The calibrators are dilutions of antibody reactive to dsDNA.Values are assigned as arbitrary anti-DNA activity units (seeLimits of the Assay).