Very Low Readings Across Plate (ELISA Development)

Question

Accepted Answer

Consistently low optical density readings across entire plate including standards Common causes: 1) Incorrect wavelengths used in plate reader 2) Insufficient development time 3) Coated plates are old and degraded 4) Capture antibody did not bind to plate 5) Buffer containing FCS used to reconstitute antibodies Solutions: 1) Check filters and reader wavelength settings 2) Increase development time 3) Coat new plates 4) Use proper ELISA plates (not tissue culture plates) 5) Dilute capture antibody in PBS without additional protein 6) Requalify reagents if FCS was used