Reliable and Reproducible MTT Assay for Cell Viability Studies

June 24, 2026

BlueskyReddit
University of Florida
Neurosurgery
Research Scientist

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Company:

Invitrogen

Product Name:

MTT Cell Proliferation Assay Reagent

Catalog Number:

M6494

MTT was prepared at 5 mg/mL in PBS, and 20 µL was added to each well containing 200 µL of medium (220 µL total volume). Cells were incubated for 4 hours at 37°C in a CO2 incubator. The medium was then removed, and 100 µL DMSO was added per well. Plates were shaken for 15 minutes before measuring absorbance at 570 nm. The assay produced consistent and reproducible cell viability measurements across experimental groups.

Experimental Design and Results Summary

Application

Cell Viability Assay (MTT)

Starting Material

Cells

Tips

Prepare MTT at 5 mg/mL in PBS. Add 20 µL MTT solution per well containing 200 µL culture medium. Protect reagent from light and include medium-only blanks for background subtraction.

Results Summary

MTT was prepared at 5 mg/mL in PBS and 20 µL was added to each well containing 200 µL of medium (220 µL total volume). Cells were incubated for 4 hours at 37°C in a CO₂ incubator. The medium was removed and 100 µL DMSO was added to each well. Plates were shaken for 15 minutes before measuring absorbance at 570 nm. The assay provided reproducible and consistent cell viability measurements.

DOI or PMID #

N/A

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Summary

The Good

Simple protocol, robust signal, reproducible results, and compatible with high-throughput screening.

The Bad

Requires a 4-hour incubation and DMSO solubilization step before analysis.

The Bottom Line

Reliable and easy-to-use MTT reagent that provides consistent cell viability data with minimal optimization.

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