Mouse Anti Nuclear Antigens (ANA/ENA) Ig's (total (A+G+M) ) ELISA Kit

cat#5210

INTENDED USE

The Mouse ANA (Anti-Nuclear Antibodies) Total Ig ELISA Kit is animmunoassay suitable for quantifying or titering total antibodyactivity (IgG, IgA and IgM) specific for extractable nuclear antigens(ENA) in serum or plasma. Other biological fluids, including tissueculture medium, may be validated for use.

PRINCIPLE OF THE TEST

The Mouse ANA Total Ig ELISA kit is based on the binding ofmouse ANA IgG, IgA and IgM in samples to ENA immobilized onthe microwells, and total ANA antibody is detected by anti-mouseIgG+IgA+IgM (H+L) specific antibody conjugated to HRP(horseradish peroxidase) enzyme. After a washing step,chromogenic substrate (TMB) is added and color is developed bythe enzymatic reaction of HRP on the substrate, which is directlyproportional to the amount of ANA Ig present in the sample.Stopping Solution is added to terminate the reaction, andabsorbance at 450nm is then measured using an ELISA microwellreader. The activity of total mouse antibody in samples iscalculated relative to mouse ANA calibrators. 

Specificity

ENA used to coat the microwells is a mixture of various extactableantigens (DNA, SSA/Ro, SSB/La, Scl70, Sm, RNP, and Jo-1); theMouse ANA assay may detect antibodies to any of these.Separate ELISA kits are available from ADI to detect antibodiesspecific to each of these individual autoantigens. The anti-MouseIgG+IgA+IgM (H+L) HRP conjugate reacts with mouse IgG, IgAand IgM class antibodies that bind to ENA on the plate. IgEantibody would not be measured above background signals.

Assay Sensitivity

The ENA coating level, HRP conjugate concentration and LowNSB Sample Diluent are optimized to differentiate ANA Ig frombackground (non-antibody) signal with mouse serum samplesdiluted 1:100.