ELISA: Best Practices and Troubleshooting

Presenter: Dr. Paul Held - Applications Laboratory Manager, BioTek Instruments

Enzyme-linked immunosorbent assay (ELISA) is an antibody-based reaction that allows the detection and quantitation of numerous analytes with the same basic assay process. The reaction uses a solid surface, typically a microplate, to immobilize the analyte. Once immobilized, unwanted materials can be removed by washing and the remaining analyte quantitated. The assay technology uses antibodies in conjunction with enzyme-catalyzed amplification to provide high specificity and high sensitivity. Here we describe the basic process steps of ELISA, best laboratory practices along with troubleshooting tips for common problems.  In addition, some of the devices and instrumentation used with this assay technology will be described.