Multiplexing AlphaLISA® type assays will increase throughput while sample expenditure and costs are minimized. The ability of the PHERAstar® FS microplate reader to perform simultaneous dual emission detection of this assay will allow users to double the information obtained from each well in the same amount of time as needed for a single emission assay.
Antibody pairs were selected for each of two protein targets. For each protein target, one of the paired antibodies was conjugated to either AlphaPLEX™ 545 nm acceptor beads or AlphaLISA® acceptor beads. The other antibody of each pair was biotinylated. A mixture containing acceptor beads and biotinylated antibodies for both target proteins was prepared and added to the reaction plates.
After laser excitation two different emission wavelengths were measured using the AlphaLISA SDE optic module. Excellent assay parameter for S/B as well as for Z' indicate the microplate readers capability to measure Alpha signals simultaneously without compromising data quality.