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Mechanistic Toxicity Assays

• Mitochondrial toxicity (using galactose in medium)
• Oxidative stress (GSH/GSSG ratio)
• Caspase 3/7 activity (apoptosis induction)

Mechanistic toxicity assays are an important part of our in vitro toxicology service portfolio, as they provide further information about the cell toxicity displayed by the study compound. All assays are with fluorescence or luminescence end-points.

Mitochondrial toxicity is assessed by measuring cytotoxicity and ATP content in the cell using galactose as carbohydrate source during short compound exposure, using glucose as comparison. Study compound with no cytotoxicity but reduced ATP production augmented by galactose, is potential mitochondrial toxicant.

Oxidative stress is assessed by measuring the ratio of an abundant antioxidant, glutathione as reduced (GSH) or oxidized (GSSG) form, from which the latter is a sign of oxidative stress. Apoptosis (programmed cell death) is activated by protein-cleaving caspases; by measuring the common caspase 3/7 pathway activation it can be assessed whether study compound induces apoptosis.