Detection of Human IgG Using a Luminescence-based Proximity Assay

This application note describes the use of the Synergy Neo Multi-Mode Microplate Reader in conjunction with the Spatial Proximity Analyte Reagent Capture Luminescence (SPARCL) assay technology to quantitate human IgG. (SPARCL) technology is a proximity dependent, chemiluminescent detection technique. In the assay, a chemiluminescent substrate (acridan) is brought into the proximity of an oxidative enzyme (horseradish peroxidase, HRP) through the specific antigen/antibody interaction. Once in close proximity adding the “trigger” reagent results in a flash luminescent signal that is proportional to the amount of analyte.