MTT reagent to assess cell viability

May 27, 2015

indian institute of chemical technology
chemical biology

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Company:

Sigma-Aldrich

Product Name:

Thiazolyl blue tetrazolium bromide

Catalog Number:

M2128

We do work on gene and protein expression changes (epigenetic modulators) in neuroglia differentiation and behaviour. This product was used to find the neuroprotective efficacy of small molecules.  MTT (Thiazolyl blue tetrazolium bromide) is an easy and effective assay to assess cell viability in different treatment conditions.

Experimental Design and Results Summary

Application

To find cell viability and identify proliferation

Starting Material

Mouse neuronal cells (N2A)

Protocol Overview

Prepare MTT reagent of 5mg/ml in 1XPBS and filter it. After incubation of cells with different treatments, add 10µl of MTT to the medium and keep in incubator (37oc) for 2h. Remove the medium by inverting the plate and let it dry. Add 100µl of DMSO mix and take the OD at 562nm. Calculate the percentage of viable cells.

Tips

Equal number of cells should be seeded. Media should be removed carefully after MTT incubation.

Results Summary

Showed neuroprotection compared to the control compound treatment.

Additional Notes

None

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Summary

The Good

Simple, less time consuming, and high-throughput assay.

The Bad

Formazan crystals come out while taking out media

The Bottom Line

It saves lot of time for high-throughput screening of compounds for cell viability and proliferation. Recommended to use.

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