Transcreener AMP2/GMP2 TR-FRET Assay

The Transcreener^® 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. Transcreener assays have been validated with major HTS instruments to determine the optimal filters and settings for maximal assay performance. Application notes with all important details are available.

The Transcreener^® AMP2/GMP2 assay is available in two fluorescent readouts, Fluorescence polarization (FP) and Time-resolved Forster-resonance-energy-transfer (TR-FRET), which provide a safe HTS-compatible alternative to cumbersome radioassay methods and are more sensitive and less subject to interference than other detection methods. Both readouts use far-red tracers to minimize compound interference and lower false-positive rates.

The Transcreener^® AMP2/GMP2 TR-FRET Assay (Prod. No. BBL-3020) is competitive immunoassay with a far-red TR-FRET readout. The assay relies on a highly specific monoclonal antibody that recognizes AMP or GMP with more than 1,000-fold selectivity over substrate nucleotides, including ATP, cAMP, or cGMP. The assay can be used with any enzyme that produces AMP or GMP, regardless of the substrate. Examples of enzymes include ubiquitin, small ubiquitin-related modifiers (SUMO), nucleic acid and protein ligases, phosphodiesterases (PDEs), and synthetases. The Transcreener^® AMP2/GMP2 FP Assay is a simple one step homogenous detection assay, and is extremely flexible with regard to substrate concentrations (0.1 to 1,000 μM). The assay provides excellent quality with a Z’ = 0.7 at low substrate conversion (typically 10-30%).