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Mitochondrial ToxGlo™ Assay Technical Manual

Instructions for Use of Product(s)
G8000, G8001

Literature # TM357

The Mitochondrial ToxGlo™ Assay is a cell-based assay method that employs a sequential-addition, multiplexed chemistry for predicting potential mitochondrial dysfunction as a result of xenobiotic exposure. The assay is based on the differential measurement of biomarkers associated with changes in cell membrane integrity and cellular ATP levels relative to vehicle-treated control cells during short exposure periods. Cell membrane integrity is first assessed by measuring the presence or absence of a distinct protease activity associated with necrosis using a fluorogenic peptide substrate (bis-AAF-R110) to measure “dead cell protease activity”. The bis-AAF-R110 Substrate cannot cross the intact membrane of live cells and therefore gives insignificant signal with viable cells, relative to non-viable cells. Next, ATP is measured by adding the ATP Detection Reagent, resulting in cell lysis and generation of a luminescent signal proportional to the amount of ATP present. The ATP Detection Reagent consists of an optimized formulation for ATP detection containing luciferin, ATPase inhibitors and thermostable Ultra-Glo™ luciferase. The two sets of data can be combined to produce profiles representative of mitochondrial dysfunction or non-mitochondrial related cytotoxic mechanisms.

Summary of Changes
The following changes were made to the 4/15 revision of this document:
1. The patent/license statements were updated.
2. The document design was updated.

Revised 4/15