MTT Assay

Cell Meter™ assay kits are a set of tools for monitoring cell viability. A variety of parameters can be used to monitor cell viability. Cell Meter™ Colorimetric MTT Cell Proliferation Kit uses MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to quantify the number of live cells. The water-soluble MTT produces water-insoluble purple formazan in metabolically active cells. The amount of formazan produced is directly proportional to the number of living cells. The absorbance increase is measured around 560 nm. Unlike other commercial MTT assays that require the tedious solubilization step to dissolve the water-insoluble formazan product, Cell Meter™ Colorimetric MTT Cell Proliferation Kit is a simple mix and read format with the minimal hands-one time. Our proprietary formulation eliminates the time-consuming solubilization step. It is the most robust MTT test for determining the number of viable cells in cell proliferation and cytotoxicity assays. The detection sensitivity is 10 times higher than the traditional MTT assays.

Example protocol

AT A GLANCE

Protocol summary

Prepare cells in a 96-well plate (100 µL/well)
Add MTT working solution (140 µL/well)
Incubate at 37 °C for 2-4 hours
Read absorbance at 560 nm

Important

Thaw all the kit components at room temperature before use.

PREPARATION OF WORKING SOLUTION

MTT working solution

Prepare the amount of MTT working solution needed by mixing 4 mL of MTT Reagent A (Component A) with 10 mL of MTT Reagent B (Component B) (2:5, v/v ratio of MTT Reagent A: B). Mix well.
Note 14 mL MTT working solution is enough for 100 tests in a 96-well plate. Prepare enough MTT working solution right before the experiment, use promptly.

SAMPLE EXPERIMENTAL PROTOCOL

Plate 1000 to 40,000 cells/well in a tissue culture microplate with clear bottom.
Add test compounds into the cells and incubate for a desired period of time (such as 24, 48 or 96 hours) in a 37 °C, 5% CO2 incubator. For blank wells (medium without the cells), add the same amount of test compounds. The suggested total volume is 100 µL for a 96-well plate, and 25 µL for a 384-well plate.
Note Each cell line should be evaluated on an individual basis to determine the optimal cell density for proliferation or cytotoxicity induction. For proliferation assays, use fewer cells; for cytotoxicity assays, use more cells to start with.
Add 140 µL/well (96-well plate) or 35 µL/well (384-well plate) of MTT working solution to each well.
Incubate the plate at 37 °C for 2-4 hours, protect from light.
Note The incubation time could be from 2 hours to overnight depending on the individual cell type and cell concentration used. Optimize the incubation time for each experiment.
Monitor the absorbance increase with an absorbance plate reader at OD = 562 nm.

Citations

View all 1 citations: Citation Explorer

Two-Pronged Intracellular Co-Delivery of Antigen and Adjuvant for Synergistic Cancer Immunotherapy
Authors: Meng, Junli and Zhang, Peisen and Chen, Qizhe and Wang, Zihua and Gu, Yuan and Ma, Jie and Li, Wang and Yang, Chen and Qiao, Yuanyuan and Hou, Yi and others,
Journal: Advanced Materials (2022): 2202168

References

View all 50 references: Citation Explorer

Novel Dental Poly (Methyl Methacrylate) Containing Phytoncide for Antifungal Effect and Inhibition of Oral Multispecies Biofilm.
Authors: Lee, Myung-Jin and Kim, Min-Ji and Oh, Sang-Hwan and Kwon, Jae-Sung
Journal: Materials (Basel, Switzerland) (2020)

Toxicity of diuron metabolites in human cells.
Authors: Mohammed, Ali Mustafa and Huovinen, Marjo and Vähäkangas, Kirsi H
Journal: Environmental toxicology and pharmacology (2020): 103409

[Comparative study on quality of decoction pieces of Saposhnikovia divaricata with different growth patterns and years and thinking of standard of decoction pieces of S. divaricata in Chinese Pharmacopoeia].
Authors: Xue, Xue and Wang, Hao and Jia, Tian-Ying and Qu, Wen-Jia and Wang, Hai-Li and Xin, Jie-Ping and Liu, Meng-Nan and Xiong, Hui and Li, Xiang-Ri
Journal: Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica (2019): 4034-4042

Evaluation of hepatotoxicity potential of a potent traditional Tibetan medicine Zuotai.
Authors: Zhou, Liang-Liang and Chen, Hai-Juan and He, Qiang-Qiang and Li, Cen and Wei, Li-Xin and Shang, Jing
Journal: Journal of ethnopharmacology (2019): 112-118

Development and validation of UPLC method for WST-1 cell viability assay and its application to MCTT HCE™ eye irritation test for colorful substances.
Authors: Joo, Kyung-Mi and Kim, Seolyeong and Koo, Ye Ji and Lee, Miri and Lee, Su-Hyun and Choi, Dalwoong and Lim, Kyung-Min
Journal: Toxicology in vitro : an international journal published in association with BIBRA (2019): 412-419

Page updated on August 13, 2024

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