Transcreener® ADP² FP Assay

The Transcreener® ADP² FP Assay from BellBrook Labs is an extremely sensitivite ADP Assay with a more sensitive antibody against ADP yielding an excellent signal at less than or equal to 10% ATP consumption for a broad range of initial ATP concentrations (0.1-1,000 μM). The result is the ability to screen low ATP Km enzymes, and to use initial velocity enzyme kinetics at or below ATP Km concentrations, which leads to accurate inhibitor potencies and the ability to use less enzyme and substrate. Ratiometric, red-shifted fluorescence polarization output minimizes signal variability and reduces compound interference.

The Transcreener® ADP² Assay is a fluorescence polarization immunoassay based on the detection of ADP by an antibody. This assay platform provides the possibility to universally interrogate all enzymes that catalyze group transfer reactions with ATP. In step one of the assay, enzymes catalyze the transfer of phosphate from ATP to a protein, peptide, lipid or small molecule resulting in the accumulation of ADP.

In step two the Transcreener® ADP² Detection Mixture, which contains an ADP Alexa633 tracer bound to an anti-ADP antibody, is added. If there is enzymatic activity resulting in necessary ADP then the bound tracer is displaced by the ADP. The free tracer rotates quickly leading to a lower polarization value. If there is no free ADP because of no enzymatic activity, the tracer is still bound to the antibody. This whole construct rotates very slowly giving a higher polarization number. Therefore, ADP production leads to a decrease in fluorescence polarization.

BMG LABTECH has also published Transcreener® ADP FP Application Notes (AN) with BellBrook Labs:

New Transcreener® ADP² FP Assay performed on BMG LABTECH´s POLARstar Omega Microplate Reader (AN183)

New Transcreener® ADP² FP Assay performed on BMG LABTECH´s PHERAstar Plus HTS Microplate Reader (AN 182)

Transcreener® ADP Fluorescence Polarization Assay Performed on the PHERAstar (AN 152)

The following BMG LABTECH microplate readers are Transcreener® ADP² FP certified from BellBrook Labs:

For more information about the Transcreener® ADP² FP Assay and other Transcreener® technologies visit:
http://www.bellbrooklabs.com