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
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.