Methyl Thiazolyl Tetrazolium or MTT Assay: A Colorimetric Assay to Measure Drug Resistance via Determination of Metabolic Activity in Cancer Cells
Published: April 30, 2023
Abstract
Source: Sciarrillo, R. et al. Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models. J. Vis. Exp. (2016)
This video describes the colorimetric assay, methyl thiazole tetrazolium (MTT) assay, to measure drug-induced cytotoxicity in cancer cells and determine their metabolic activity. This assay is based on the metabolic conversion of tetrazolium salts into colored products, reflecting the mitochondrial activity of cells.
Protocol
1. MTT Assay for Leukemic Cells Prepare in advance the MTT solution: Dissolve 500 mg of MTT formazan in 10 mL PBS and stir (protected from light) with a magnetic stirrer for approximately 1 h. Sterilize the solution with a 0.22 µm filter. NOTE: The solution can be stored in 10 mL aliquots at −20 °C. Protect from direct light after thawing. Prepare in advance the acidified isopropanol: Add 50 mL of 2 M HCl to 2.5 L of isopropanol. <stron…
Açıklamalar
The authors have nothing to disclose.
Materials
| CCRF-CEM | ATCC, Manassas, VA, USA | ATCC CCL-119 | |
| RPMI-1640 | Gibco, Carlsbad, CA, USA | 11875093 | |
| Fetal bovine and calf serum | Greiner Bio-One, Frickenhausen, Germany | 758093 | |
| penicillin G streptomycin sulphate | Gibco, Carlsbad, CA, USA | 15140122 | |
| Tris(hydroxymethyl)-aminomethane | Sigma Aldrich | 252859 | |
| CELLSTAR Cell Culture Flasks 25 centrimeter square | Greiner Bio-One, Frickenhausen, Germany | 82051-074 | |
| MTT formazan | Sigma Aldrich | M2003 | |
| Anthos-Elisa-reader 2001 | Labtec, Heerhugowaard, Netherlands | UV-Vis 96-well plate spectrophotometer | |
| Greiner CELLSTAR 96 well plates | tes Greiner/Sigma | M0812-100EA | |
| Phosphate Buffered Saline (NaCl 0.9%) | B.Braun Melsungen AG, Germany | 362 3140 | |
| Trypsin/EDTA Solution 100 ml | Lonza, Basel, Switzerland | CC-5012 |
Etiketler
Bu Makaleden Alıntı Yapın
Methyl Thiazolyl Tetrazolium or MTT Assay: A Colorimetric Assay to Measure Drug Resistance via Determination of Metabolic Activity in Cancer Cells. J. Vis. Exp. (Pending Publication), e20491, doi: (2023).