MTT Assay
Mitochondrial dehydrogenase activity measured via solubilised formazan.
To assess cell viability, proliferation and cytotoxicity.
Similar to MTS, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) is reduced by mitochondrial dehydrogenases in viable cells to an insoluble purple formazan crystal. Unlike MTS, this formazan must be solubilised before measurement.
Standard protocol
Protocol (typical)
- 1
Seed cells in a 96-well plate and incubate.
- 2
Treat cells with test agents.
- 3
Add MTT solution (e.g., 5 mg/mL in PBS, 10–20 μL per well).
- 4
Incubate at 37 °C for 2–4 hours.
- 5
Remove the medium and carefully aspirate.
- 6
Add solubilisation solution (e.g., DMSO, isopropanol or acidified SDS) to dissolve formazan crystals.
- 7
Shake gently to ensure complete dissolution.
- 8
Measure absorbance at 570 nm (reference ~630–690 nm).
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