UGT Activity Assays

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 an...

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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. The UGT-Glo™ Screening Systems contain the above reagents as well as the respective UGT isoforms and control membranes.

Features - Benefits

• Speed: The luminescent format eliminates the need for time-consuming analyses such as HPLC and LC/MS.
• Simplified Method: The simple "add and read" protocol makes the assay amenable to higher throughput screening in multiwell plates.
• Sensitive: Allows researchers to use less enzyme and scale down reaction volumes, which saves on reagent costs.

Applications

• Screen drugs and new chemical entities for their capacity to modulate UGT activity in native or recombinant fractions.
• Measure recombinant UGT activities in membrane fractions from heterologous expression systems, such as insect cells and E. coli.

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