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The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay is a widely used method to assess cell metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. The assay is based on reducing the tetrazolium salt MTT to formazan by mitochondrial enzymes in living cells. This chapter describes the detailed protocol for performing the MTT assay, including preparation of reagents, cell plating, MTT incubation, and formazan solubilization. Key considerations and troubleshooting tips are also discussed. Additionally, available tools for calculating IC50 are also indicated.

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From Protocol to Analysis: Performing and Interpreting the MTT Assay

  • 章节

  • 发表日期：

    2026年

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  • Andreia Rego,
  • José Furtado,
  • José A. Silva,
  • Paula Alvito,
  • Carla Motta

摘要

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay is a widely used method to assess cell metabolic activity as an indicator of cell viability, proliferation, and cytotoxicity. The assay is based on reducing the tetrazolium salt MTT to formazan by mitochondrial enzymes in living cells. This chapter describes the detailed protocol for performing the MTT assay, including preparation of reagents, cell plating, MTT incubation, and formazan solubilization. Key considerations and troubleshooting tips are also discussed. Additionally, available tools for calculating IC50 are also indicated.