The UGT-Glo™ Assay provides a luminescent method for measuring UDP glucuronosyltransferase (UGT) activity. The UGT-Glo™ Assay is designed to measure UGT activity from a variety of sources, such as microsomes containing recombinantly expressed enzymes or microsomal preparations derived from mammalian tissues, and to test the effects of various chemicals on UGT activity.
The assay involves incubating UGT with a proluciferin substrate; a portion of the substrate gets conjugated with UDP, while the remainder is unmodified. Upon the addition of d-Cysteine, the unconjugated proluciferin is converted into luciferin and, in a coupled reaction with luciferase/luciferin, is converted into light. Conjugated proluciferin remains intact and does not contribute to the luminescence. Thus, the signal generated is inversely correlated with UGT activity present in the sample.
The UGT-Glo™ Assay contains two proluciferin substrates: the UGT Multienzyme Substrate, which is compatible with a wide range of UGTs, and the UGT1A4 Substrate, which reacts specifically with UGT1A4. The kit also contains Luciferin Detection Reagent and Reconstitution Buffer, UGT Buffer, d-Cysteine and UDPGA.