Calculator
Cell Meter™ Colorimetric WST-8 Cell Quantification Kit
Example protocol
AT A GLANCE
Prepare cells in a 96-well plate (100 µL/well).
Add 10 µL of WST-8™ Solution to each well.
Incubate at 37°C for 1 - 4 hours.
Monitor absorbance at OD = 460 nm.
The WST-8™ Solution can be stable for up to one year if stored at 4°C and protected from light. Store it at <-20°C for longer storage.
CELL PREPARATION
For guidelines on cell sample preparation, please visit:
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Plate 5000 to 10,000 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 or 50 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 10 µL/well (96-well plate) or 5 µL/well (384-well plate) of WST-8™ Solution to each well.
Incubate the plate at 37°C for 1 - 4 hours, protect from light.
Note: The incubation time could be from 30 minutes 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 =460 nm.
Prepare cell culture in a tissue culture microplate with a clear bottom. The suggested total volume is 100 µL for a 96-well plate or 50 µL for a 384-well plate.
Note: We used serially diluted HeLa and Jurkat cell suspension in a clear bottom 96-well plate for the assay.
Add 10 µL/well (96-well plate) or 5 µL/well (384-well plate) of WST-8™ Solution to each well.
Incubate the plate at 37°C for 1 - 4 hours, protect from light.
Note: The incubation time could be from 30 minutes 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 = 460 nm.
Citations
Authors: Finburgh, Emma N and Mauduit, Olivier and Noguchi, Takako and Bu, Jennifer J and Abbas, Anser A and Hakim, Dominic F and Bellusci, Saverio and Meech, Robyn and Makarenkova, Helen P and Afshari, Natalie A
Journal: Investigative Ophthalmology \& Visual Science (2023): 21--21
Authors: Peng, Lilei and Ming, Yang and Zhang, Ling and Zhou, Jie and Xiang, Wei and Zeng, Shan and He, Haiping and Chen, Ligang
Journal: The FASEB Journal (2020): 5128--5143
Authors: Gao, Yong and Sun, Laisheng and Wu, Zicheng and Xuan, Chengmin and Zhang, Junxia and You, Yongping and Chen, Xincheng
Journal: Molecular Medicine Reports (2017)
References
Authors: Tiwari, K.; Wavdhane, M.; Haque, S.; Govender, T.; Kruger, H. G.; Mishra, M. K.; Ch and ra, R.; Tiwari, D.
Journal: Pharm Biol (2015): 849-54
Authors: Tsukatani, T.; Suenaga, H.; Shiga, M.; Noguchi, K.; Ishiyama, M.; Ezoe, T.; Matsumoto, K.
Journal: J Microbiol Methods (2012): 160-6
Authors: Stoddart, M. J.
Journal: Methods Mol Biol (2011): 21-5
Authors: Brady, A. J.; Kearney, P.; Tunney, M. M., Comparative evaluation of 2,3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT) and 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium
Journal: J Microbiol Methods (2007): 305-11
Authors: Arai, M.; Kosuge, K.; Kawamoto, F.; Matsuoka, H.
Journal: Acta Med Okayama (2006): 127-34
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