Cell viability (MTT Assay)

Solutions

MTT-Solution:

5mg/mL (≙12mM) (toxic and mutagenic) solved in PBS, sterile filtered with 0,2 µM filter

Formazan solving solution: isopropanol

Procedure

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Coat plate before usage

For 96 well plates: use 1 - 2x104 cells in 100 µL medium per well and treat the cells according to your experimental setup.

The cell number above was optimized for 1 day treatment; for longer incubation periods use fewer cells accordingly.

• After treatment period add 10 µL of the MTT-PBS solution
• Incubate at 37°C for 1-2h.
• Remove the medium completely via inversion (if you have non adherent cells or if the adherence is weak centrifuge the cells beforehand for 3 min at 800 rpm).
• Add 100 µL isopropanol to dissolve to formazan cristals
• Incubate overnight at room temperature
• Check in the microscope for the solubilisation of the formazan crystals – if they are still visible incubate longer
• Measure the absorbance at 550 nm in plate reader

Note: cell number was optimized for HEK cells - if you use other cells you may have to alter the conditions accordingly.