Epstein Barr Virus Nuclear Antigen IgG ELISA

The Epstein-Barr Virus (EBNA-1) IgG ELISA Kit is a solid-phase enzyme-linked immunosorbentassay (ELISA). Microtiter wells as a solid phase are coated with recombinant EBV nuclear antigentype 1. Diluted samples and ready-for-use controls are pipetted into these wells. Duringincubation, EBNA-1-specific antibodies of positive samples and controls are bound to theimmobilized antigens. After a washing step to remove unbound sample and control material,horseradish peroxidase-conjugated anti-human IgG antibodies are dispensed into the wells. Duringa second incubation, this anti-IgG conjugate binds specifically to IgG antibodies resulting in theformation of enzyme-linked immune complexes. After a second washing step to remove unboundconjugate, the immune complexes formed (in case of presence of antigen) are detected byincubation with TMB substrate and development of a blue color. The blue color turns into yellow bystopping the enzymatic indicator reaction with sulfuric acid. The intensity of this color is directlyproportional to the amount of EBNA-1-specific IggG antibody in the sample. Absorbance at 450 nmis read using an ELISA microtiter plate reader.