We use cookies to provide visitors of our website with the best possible experience. To learn more how we use cookies or how to block cookies, please visit our cookie policy.

Resources - Scientific Posters

Volume Scaling of the Transcreener® ADP2 FP Assay on the BioTek Synergy™ 2 and 4 Multi-Mode Microplate Readers Allows For Easy Transition From Assay Development to High-Throughput Screening

Download

February 02, 2009

 

Authors: Tracy Worzella, Brad Larson, Karen Kleman-Leyer, BellBrook Labs; Xavier Amouretti, Peter Banks, BioTek Instruments

 
BellBrook Labs
 

Overview

 

We present data showing the validation of BellBrook Labs’ Transcreener® ADP2 FP Assay on the BioTek Synergy™ 2 and Synergy™ 4 plate readers. The Transcreener ADP2 Assay is a universal, second generation far red fluorescence polarization assay that detects the ADP nucleotide by-product from any ATP-utilizing enzyme reaction. The assay is amenable to volume scaling, as shown by data generated in 96-well and 384-well microplate formats. Data generated with a standard curve in both microplate formats resulted in Z´ values ≥ 0.6. Further, the 96-well assay using the standard PMT and the 384-well assay using the red-shifted PMT meet BellBrook Labs' Instrument Validation Program criteria, which include Z´ > 0.7 with Δ mP shift ≥ 95 at 10% conversion of 10 μM ATP to ADP. Further enzyme studies demonstrate that 96- and 384-well plate formats yield excellent Z´-factor results with Protein Kinase A (PKA) at low % ATP conversion. Known PKA inhibitors show comparable potency in both 96- and 384-well plate formats, and between different Synergy plate readers. Although the standard PMT Synergy does not meet validation criteria in 384-well plate format, comparable inhibitor potency results are obtained between the standard and red-shifted PMT instruments.


 

link