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Promega Corporation

Determine Mechanism of Toxicity with Mitochondrial ToxGlo™ Assay

Select an area of the image to view representative toxicity profiles

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No Mitotoxicity

No Mitotoxicity

No changes in ATP or membrane integrity = No Mitotoxicity

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.

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Mitotoxicity With No Necrosis

Mitotoxicity - No Necrosis

Reduction in ATP with no necrosis = Mitochondrial toxicity

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.

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Mitotoxicity With Necrosis

Mitotoxicity - With Necrosis

Ablation of ATP leads to dose-dependent membrane integrity changes (necrosis) = Mitochondrial toxicity

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.

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Primary Necrosis

Primary Necrosis

Reduction in ATP with commensurate membrane integrity changes = Primary Necrosis

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.

Assay Overview

Single point assays of viability or cytotoxicity are not sufficient to determine mechanism of toxicity. The Mitochondrial ToxGlo™ Assay brings you the predictive data you need to distinguish mitochondrial dysfunction from other cytotoxic effects.

  • Predictive for mitochondrial toxicity
  • Results in under an hour
  • Easy automation
  • Simple detection with a standard plate reader

Assay Principle

The Mitochondrial ToxGlo™ Assay is based on the differential measurement of biomarkers associated with changes in cell membrane integrity and cellular ATP levels relative to vehicle-treated controls. The assay is performed in a single-well, with bioluminescent and fluorescent readouts.

  • Bioluminescent signal is proportional to ATP concentration
  • Fluorescent signal indicates loss of membrane integrity
  • The two sets of data are combined to produce profiles representative of mitochondrial dysfunction or other cytotoxic mechanisms.

Simple, "Add-Mix-Measure" Protocol

Simple add-mix-measure protocol is easy to implement and fast. No parallel processing in glucose-containing medium.

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