Anti-Liver-Kidney Microsomes-1 IgG ELISA (Anti-LKM-1 IgG ELISA)

Serum samples diluted 1:101 are incubated in the microplates coated with the specific antigen. Antibodies, if present in the samples, bind to the antigen. The unbound fraction is washed off in the following step. Afterwards, anti-human immunoglobulins conjugated to horseradish peroxidase (conjugate) are incubated and react with the antigen-antibody complex of the samples in the microplates. Unbound conjugate is washed off in the following step. Addition of TMB substrate generates an enzymatic colorimetric (blue) reaction, which is stopped by diluted acid (color changes to yellow). The intensity of color formation from the chromogen is a function of the amount of conjugate bound to the antigen-antibody complex and this is proportional to the initial concentration of the respective antibodies in the sample.