Outstanding performance of THUNDER™ on BioTek®’s Synergy™ Neo2 multimode microplate reader

Bioauxilium has initiated strategic collaborations with leading developers of microplate detection instruments to demonstrate the compatibility of its increasingly popular THUNDER™ TR-FRET assay platform with select high-value multimode microplate readers under real assay conditions.

Agilent BioTek® has recently evaluated the performance of the THUNDER™ Phospho-AKT pan (S473) TR-FRET Cell Signaling Assay Kit on the Synergy™ Neo2 multi-mode microplate reader. The results, summarized in an Application Note, showed an outstanding assay performance on this instrument.

Z’-factor determination in NIH3T3 cells (10,000 cells/well) incubated with 10 nM PDGF-AA for 15 minutes at 37 °C and then lysed.

Highlights

• The THUNDER™ Phospho-AKT pan (S473) Assay Kit was tested in a 384-well format using the 2-plate assay protocol.
• NIH/3T3 cells plated at 10,000 cells per well were treated with increasing concentrations of either PDGF or wortmannin prior to stimulation with 10 nM PDGF.
• Assays were read on the Synergy Neo2 equipped with a 377 nm laser excitation source.
• PDGF and wortmannin concentration-responses curves generated signal-to-background ratios of 9 and EC50 and IC50 values of 1.5 nM and 56 nM, respectively.
• A manual Z’-factor experiment generated a value of 0.72, demonstrating the robustness of the cellular assay.
• The assay detected as few as 2,500 PDGF-treated cells per well against a no-cell control with a Z’-factor of 0.47.
• Agilent BioTek’s Synergy Neo2 is therefore certified for use with THUNDER™.