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ADME-Tox Assays

Luminogenic substrates bring the advantages of bioluminescence to the enzymology researcher studying the absorption, distribution, metabolism or excretion of molecules. Our selection of bioluminescent enzyme substrates for proteases and metabolic enzymes such as CYPs ,  MAO and Pgp  provide powerful tools for ADME-tox screens.

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ADME-Tox Assay Basics

ADME assays, used to assess absorption, distribution, metabolism, and excretion, are critical for evaluation of drug candidates and other pharmaceuticals. Promega assays relevant to ADME screening detect cytochrome P450, p-glycoprotein, and monoamine oxidase activities, and total glutathione as an indicator of oxidative stress.

The majority of small-molecule drugs are metabolized by cytochromes P450 (CYP450) enzymes. CYP450-mediated metabolism influences clearance rates of drugs, their toxicity and their interactions with coadministered drugs. Drug discovery researchers need to determine how new drug entities are metabolized by CYP450s and to what extent they may alter CYP450 activity. P450-Glo™ CYP450 Assays provide a homogeneous, luminescent method for measuring cytochrome P450 activity. They are designed for measuring the activities of P450s from recombinant and native sources and for testing the effects of analytes such as drugs and new chemical entities on P450 activities in a multiwell format. P450-Glo™ Luminescent Assays exhibit exquisite sensitivity, low background signals and broad dynamic range.

Monoamine oxidases (MAOs) are flavoenzymes located in the outer membrane of mitochondria that catalyze the oxidative deamination of a number of biogenic and xenobiotic amines. MAOs can oxidize neurotransmitters and produce toxic H2O2. The ability to measure MAO activity and the effects of test compounds on that activity is critical for selecting specific MAO inhibitors, discovering potential drug-drug interactions and assessing the MAO-catalyzed detoxification or bioactivation of various target compounds. The MAO-Glo™ Assay is a simple, homogeneous and robust bioluminescent assay for the rapid and sensitive detection of MAO activity.

P-glycoprotein (Pgp) is an integral plasma membrane protein that functions as an ATP-dependent drug efflux pump and plays an important role in multidrug resistance and certain adverse drug-drug interactions. Compounds that interact with Pgp can be identified as stimulators or inhibitors of its ATPase activity. The Pgp-Glo™ Assay is a luminescent assays that detects the effects of compounds on recombinant human Pgp in a cell membrane fraction. Pgp-dependent decreases in luminescence reflect ATP consumption by Pgp; thus the greater the decrease in signal, the higher the Pgp activity. 

The GSH/GSSG-Glo™ Assay detects and quantifies total glutathione (GSH + GSSG), GSSG and GSH-to-GSSG ratios in cultured cells. Changes in GSH and GSSG levels are important when assessing toxicological responses and an indicator of oxidative stress, potentially leading to apoptosis or cell death. The GSH and GSSG detection assay provides a simple, rapid multiwell-plate format where stable luminescent signals are correlated with either the total GSH or the GSSG concentration of a sample directly in culture wells.