384-Well Microplate Time Resolved Fluorescence (TRF) Immunoassay Using A Matched Antibody Pair Kit And The PHERAstar® FSX

By Jamie Campbell, Director of Custom Solutions EMEA – abcam, and Oliver Carney, BMG LABTECH

This content is brought to you by Abcam, a Danaher Operating Company.

Many commercially available antibody pairs are designed for use in sandwich ELISA assays. They provide the flexibility to scale experiments down to 384-well and 1536-well formats, accommodating high-throughput needs. Matched antibody pairs (MAPs) from Abcam are well-characterized, incorporate recombinant antibodies, and are highly sensitive reagents, covering a broad range of targets. A typical MAP includes an unlabelled capture antibody, a biotinylated detection antibody, and a protein standard.

Alternative measurement methods and secondary antibody labels can improve assay performance. For instance, Time-Resolved Fluorescence (TRF) detection employs long-lifetime fluorophores called lanthanides, which emit light over a longer duration after excitation (microseconds instead of nanoseconds). This minimizes background noise by delaying the measurement until after the background signal has dissipated. Using non-visible light spectra for assay signal measurement can enhance dynamic range and is not limited by saturation. Explore how Abcam’s MAP kits can be employed in a 384-well TRF immunoassay (DELFIA® TRF – Perkin Elmer) and analyzed using the PHERAstar® FSX to detect IRAK4, a protein kinase involved in immune response cell signaling.