Transcreener(R) ADP2 FP Assay

The Transcreener(R) HTS Assay platform overcomes the need for time-consuming, one-off assay development for individual members within a group transfer enzyme family by utilizing a single set of assay reagents that detect an invariant product. The generic nature of the Transcreener HTS Assay platform eliminates delays involved in assay development for new HTS targets, and greatly simplifies compound and inhibitor profiling across multiple target families. The Transcreener ADP2 FP Assay is a far-red, competitive fluorescence polarization (FP) assay based on the detection of ADP and therefore is compatible with any enzyme class that produces ADP, including protein, lipid, and carbohydrate kinases, ATPases, DNA helicases, carboxylases and glutamine synthetase. The Transcreener ADP2 Assay is a simple one step homogenous detection assay, and is extremely flexible with regard to ATP concentration (0.1 to 1,000 µM ATP). The assay provides excellent signal at low substrate conversion, with a Z' > 0.7 and > 85 polarization shift (mP) at 10% ATP conversion using 1 µM ATP. The Transcreener ADP2 FP Assay was developed to follow the progress of any enzyme that produces ADP. The Transcreener ADP Detection Mixture comprises an ADP Alexa633 Tracer bound to an ADP2 Antibody. The tracer is displaced by ADP, the invariant product generated during the enzyme reaction The displaced tracer freely rotates leading to a decrease in fluorescence polarization. The assay uses a far red tracer to minimize interference from fluorescent compounds and light scattering.