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pp 1-9 | Cite as

Detection of Total UDP-Glucuronosyltransferase (UGT) Activity in Melanoma Cells

  • Ryan W. Dellinger
  • Frank L. MeyskensJr.
Protocol
Part of the Methods in Molecular Biology book series

Abstract

The UDP-glucuronosyltransferases (UGTs) are integrally involved in the clearance of a wide range of drugs used to combat human diseases. UGT expression levels and activity can be induced by drug addition to cells and has been proposed as a potential intratumoral drug resistance mechanism. Traditional methods of assaying UGT activity are drug-centric and require HPLCs with multiple detectors (dependent on individual drug). Here, we describe a generalized method to detect total UGT activity (intrinsic or induced) via the UGT-Glo assay which utilizes a general UGT substrate with luminescence as the readout eliminating the need for multiple HPLC detectors to detect total UGT activity in a given sample. The method detailed here can be applied for any UGT containing sample, allowing for the efficient detection of total UGT activity to be the functional endpoint using a plate reader. In this manner, global changes in UGT activity can be monitored in response to a wide variety of stimuli.

Keywords:

UDP-glucuronosyltransferase Glucuronidase activity UGT-Glo Melanoma Drug resistance 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Chao Family Comprehensive Cancer Center and Departments of Medicine (RWD, FLM), Biological Chemistry (FLM), Epidemiology (RWD) and Public Health (FLM)University of California IrvineIrvineUSA

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