Assay Information
Purified antigens (SSA/Ro, SSB/La, RNP 70, Sm, RNP/Sm, Scl70, centromere B and Jo1) are bound to microwells. Antibodies to these antigens, if present in diluted serum, bind in the microwells. Washing of the microwells removes unbound serum antibodies. Horseradish peroxidase (HRP) conjugated antihuman IgG immunologically binds to the bound patient antibodies forming a conjugate/antibody/antigen complex. Washing of the microwells removes unbound conjugate. An enzyme substrate in the presence of bound conjugate hydrolyzes to form a blue color. The addition of an acid stops the reaction forming a yellow endproduct. The intensity of this yellow color is measured photometrically at 450nm.