BellBrook Labs scientists have developed a new universal UDP detection assay for the measurement of Glycosyltransferase activity. The Transcreener UDP2 FI Assay extends the Transcreener platform for UDP detection by utilizing a simple fluorescence intensity (FI) output. It can be used on fluorescence readers typically found in academic and therapeutic research laboratories, as well as more complex multimode plate readers used in core facilities and HTS laboratories.
The assay employs a competitive FI method, which was designed specifically for high-throughput screening (HTS), with a single-addition, mix-and-read format. Enzyme activity is signaled by an increase in fluorescence intensity as the bound tracer is displaced from the Transcreener UDP2 Antibody-Quencher. The AlexaFluor® 594 tracer uses the same plate reader settings as commonly used Texas Red. FI can provide faster read times than FP or TR-FRET. The assay is a simple single-step mix-and-read format enabling the use of unmodified native substrate concentrations of 1 – 100 µM.
The Transcreener UDP2 FI Assay is a universal biochemical HTS assay for enzymes that produce UDP, including:
- Glycogen, Cellulose, Lactose, and Hyaluronan Synthases
The Transcreener® UDP2 FI Assay provides the following benefits:
- Accommodates UDP concentrations ranging from 1 µM to 100 µM.
- Excellent data quality (Z’ ≥ 0.7) and signal at low substrate conversion using 1 µM UDP.
- Overcomes the need for time-consuming, one-off assay development for individual members within a group transfer enzyme family by using a single set of assay reagents that detect an invariant product.
- Red tracer further minimizes interference from fluorescent compounds and light scattering.