Indirect Elisa Diagram. For accurate quantitative results always compare signal of unknown samples against those of a standard curve. The primary antibody is incubated with the antigen followed by the incubation with the secondary antibody.
In 1971 elisa was introduced by peter perlmann and eva engvall at stockholm university in sweden. In an indirect elisa we quantify antigen specific antibody rather than antigen. Next an enzyme labeled secondary antibody is bound to the first antibody.
Standards duplicates or triplicates and blank must be run with each plate to ensure accuracy.
See direct elisa protocol buffers and reagents. Different strategies for both capture and detection are used in elisa. In an indirect elisa we quantify antigen specific antibody rather than antigen. Next an enzyme labeled secondary antibody is bound to the first antibody.
