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ELISA Technology

ELISA (ENZYME-LINKED IMMUNOSORBENT ASSAY) is a widely used biochemical technique for the detection of an antigen in a sample. The sandwich ELISA utilizes two antigen specific antibodies; one capture antibody bound to a solid phase and one enzyme-linked detection antibody. Direct enzyme conjugation of the detection antibody ensures an easy-to-use and sensitive assay with minimal background signal.

Sandwich ELISA sequence assay 

- antigen and detection antibody is added stepwise

In a sequence assay, sample is applied to an antibody-coated microtiter plate well. The capture antibody binds to the antigen in the sample. Unbound compounds are removed by a washing procedure before the enzyme-linked detection antibody is added to the well. The detection antibody binds to a second epitope (immunogenic part) on the antigen. Unbound antibodies are removed by washing before theaddition of a substrate, which is converted by the enzyme to a chromogenic signal. The enzyme reaction is stopped and the result is monitored spectrophotometrically. The more antigen in the sample, the stronger the signal. 

Sandwich ELISA simultaneous assay 

- antigen and detection antibody is added simultaneously

The sandwich ELISA may also be constructed as a simultaneous assay where the sample and detection antibody is added in the same step.