Mitochondrial ToxGlo Assay
Simple, fast determination of mitochondrial toxicity
Distinguish Primary Mitochondrial Dysfunction from Secondary Cytotoxic Events
- Predictive for mitochondrial toxicity
- Add-mix-measure protocol
- Results in under one hour
- Flexible and easily automated
- Detect with a standard multimode plate reader
Assay Principal
Cell-Based, multiplexed method measures ATP (a proximal measure of mitochondrial function) in conjunction with a membrane integrity biomarker (protease) to distinguish primary mitochondrial dysfunction from secondary cytotoxic events directly in the same sample well.
Protocol Overview
Simple add-mix-measure protocol is easy to implement and fast. No parallel processing in glucose-containing medium.
Toxicity Profiles
Produces profiles that are predictive for mitochondrial toxicity and discernable from other non-mitotoxic mechanisms of cell death.
K562 Cells (10,000/well) in 96-well plates were exposed to serial dilutions of test compound for 2 hours in glucose-free (galactose-supplemented) RPMI medium.
Applications
Mitochondrial ToxGlo™ Assay demonstrates appropriate responses with known Ox/Phos poisons and iCell™ Cardiomyocytes derived from human pluripotent stem cells.
Cells were exposed to treatments for 2 hours in galactose-dosing medium.
Automation Compatible
Easily scalable and automatable from 96- to LV384-well plate formats with samewell multiplexing to differentiate mitochondrial toxicity from non-mitotoxic mechanisms of cell death.
K562 cells in LV384-well plates were treated with toxicants for 2 hours in either galactose-supplemented or glucose-supplemented medium. Cells grown and/or treated in galactose-supplemented medium are poised for oxidative phosphorylation making them more sensitive to mitochondrial toxins and, detectable in this same-well multiplex assay.