MTT Assay

MTT assay is the colorimetric assay to observe the cellular metabolic activity.

Principles of MTT assay:

The MTT (3-(4, 5-dimethyl thiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay provides a readout of cell viability & growth that works by measuring cell metabolic activity. The yellow tetrazolium MTT (3-(4, 5-dimethyl thiazolyl-2)-2, 5-diphenyltetrazolium bromide) is reduced by the metabolically active cells, by the action of dehydrogenase enzymes, to produce reducing equivalents such as NADH and NADPH. The resulting intracellular purple formazan can be solubilized and quantified by spectrophotometric means at 570nm.

MTT is sensitive to light. This assay should be performed in the dark.

The utility of this assay:

This assay measures, cellular metabolic activity as an indicator of cell viability, proliferation and cytotoxicity.

MTT protocol:

1. Use 24 well plates and seed cells in each well
2. Check the cell confluency
3. Add the desired amount of required drugs
4. Incubate for 24 hours
5. Add the required amount of MTT media solution per well
6. Incubate for 4-6 hours at 37°C +5% CO₂
7. Carefully remove media with a pipette without disturbing crystals
8. Add the required amount of DMSO per well to solubilize the crystals and pipette well to mix
9. Incubate 15-30 minutes at room temperature or at 37°C
10. Use a microplate reader to read out results at an absorbance of 570 nm

Must read: Useful and important Numbers for Cell culture

Social media link:

Facebook link: https://www.facebook.com/share/p/czwDpgKbcd1h7PPn/?mibextid=oFDknk

Instagram link: