Learn how to optimize HTRF settings in the SpectraMax microplate reader.
Sponsored Content by Molecular Devices UK LtdReviewed by Aimee MolineuxSep 24 2025 HTRF assays from Cisbio have advanced to cover diverse areas, including G protein-coupled receptors, kinases and cell signaling, epigenetics, and biomarkers.
A broad range of toolbox reagents and kits is offered to support studies on protein-protein interactions, nuclear receptor assays, receptor dimerization, ligand binding, receptor internalization, enzyme assays, and other applications. HTRF uses two fluorophores, a donor and an acceptor, to label proteins or other targeted biomolecules. When these biomolecules interact, their proximity enables fluorescence resonance energy transfer from the donor to the acceptor fluorophore. The use of long-lifetime fluorescent lanthanide cryptates as donors enables time-resolved detection of HTRF assays, reducing short-lived background fluorescence from compounds and other materials. Detecting time-resolved FRET using a microplate reader provides a readout of biomolecular binding. A microplate reader must meet Cisbio’s performance standards to achieve reliable HTRF results. Key factors include the reader’s optics, energy source, detectors, and detection settings. Cisbio readers use tailored components for HTRF detection, such as a dedicated detection cartridge for the SpectraMax i3x and Paradigm readers, and a TRF module with verified filters for the SpectraMax iD5. Cisbio’s HTRF certification program ensures that readers displaying the “HTRF compatible” logo fulfill the necessary requirements for the best HTRF performance. This article outlines the optimized settings verified by Cisbio for Molecular Devices HTRF-compatible microplate readers.Figure 1. HTRF-compatible SpectraMax Multi-Mode Readers. SpectraMax i3/i3x and Paradigm readers can be equipped with the HTRF detection cartridge. The SpectraMax iD5 reader uses the HTRF detection system, which includes an Enhanced TRF Module and filters for excitation and emission. Image Credit: Molecular Devices UK Ltd About the reader setup recommendations HTRF assays are offered in three distinct donor/acceptor fluorophore combinations, each necessitating specific reader configurations to ensure optimal results:SpectraMax Paradigm reader Source: Molecular Devices UK Ltd SpectraMax M5e and FlexStation 3 readers Source: Molecular Devices UK Ltd Conclusion Using its standard certification methods, Cisbio evaluated SpectraMax multi-mode microplate readers with time-resolved fluorescence detection. The readers exceeded HTRF performance requirements and are supplied with optimized cartridges and filters to support the full range of assays, including Tag-lite. Acknowledgements Produced from materials originally authored by Cathy Olsen, Sr. Applications Scientist at Molecular Devices. About Molecular Devices UK Ltd Molecular Devices is one of the world’s leading providers of high-performance bioanalytical measurement systems, software, and consumables for life science research, pharmaceutical, and biotherapeutic development. Included within a broad product portfolio are platforms for high-throughput screening, genomic and cellular analysis, colony selection, and microplate detection. These leading-edge products enable scientists to improve productivity and effectiveness, ultimately accelerating research and the discovery of new therapeutics. Molecular Devices is committed to the continual development of innovative solutions for life science applications. The company is headquartered in Silicon Valley, California, with offices around the globe. For more information, please visit www.moleculardevices.com. Sponsored Content Policy: News-Medical.net publishes articles and related content that may be derived from sources where we have existing commercial relationships, provided such content adds value to the core editorial ethos of News-Medical.Net which is to educate and inform site visitors interested in medical research, science, medical devices and treatments.
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