Nonradioactive Receptor Binding Assays

The use of biotin labeled ligands in receptor binding assays will be used as a substitute for radioHigh affinity biotinylated ligands are used in immunochemistry studies but are not generally employebinding experiments. Biotinylated ligands also have high affinities for the proteins avidin and streproteins can be coupled with various enzymes which catalyze spectrophotometric, fluorometric, or chedetection in microliter plates. Using standard receptor binding protocols, biotinylated ligands canappropriate receptor and possible competitors for the receptor. Free or receptor bound biotinylatedby binding to the avidin or st reptavidin enzyme conjugates. This mechanism makes it possible for onbiotinylated ligand to be responsible for the generation of many molecules of product, providing aninitial signal. Several strategies for selected receptor binding assays, utilizing biotinylated ligaassay format assays will be examined. The knowledge gained in these investigations should aid in theadditional high throughput nonradioactive receptor binding assays.