Wiebke Stahlschmidt*, Christian Oberdanner*, Michael Fejtl*, Sandrine Cabirol**, Delphine Jaga**, François Degorce**
* Tecan Austria GmbH, Untersbergstr. 1a, Grödig, Austria; ** Cisbio Bioassay, Parc Marcel Boiteux, BP 84175, 30200 Codolet, France
HTRF (homogeneous time-resolved fluorescence) is a time-resolved fluorescence resonance energy transfer (TR-FRET)-based assay system from Cisbio Bioassays for the analysis of various molecular interactions and binding studies. Based on energy transfer between a long-lived europium or terbium cryptate donor and a variety of red or green acceptors, the assay combines the advantages of FRET and TRF. HTRF assays place high sensitivity demands on multimode readers. Tecan’s latest multimode reader, the Spark 10M, offers a number of features to meet these demands and ensure optimal assay performance, including the unique Fusion Optics. Fusion Optics combine the sensitivity of filters and the flexibility of monochromators (MCRs) in one instrument, offering a choice of entirely filter- or MCR-based measurements, or simultaneous use of a combination of the two. Automated Z-position optimization and an integrated dichroic mirror that is perfectly suited to HTRF measurements further increase the Spark 10M’s outstanding sensitivity. This poster describes the results of an in-depth validation of the Spark 10M reader for HTRF measurements using different Cisbio Bioassay kits, including the HTRF reader control kit designed for performance validation of multimode readers, the Human TNFα assay kit, the cAMP HiRange assay kit and the IP-One Tb assay kit.