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Cell Meter™ Colorimetric WST-8 Cell Quantification Kit

Cell number was determined with Cell Meter™ Colorimetric WST-8 Cell Quantification Kit. HeLa cells at 0 to 10,000 cells/well/100 µL were added in a clear bottom 96-well plate. The absorbance was measured at 460 nm using a SpectraMax reader (Molecular Devices).
Cell number was determined with Cell Meter™ Colorimetric WST-8 Cell Quantification Kit. HeLa cells at 0 to 10,000 cells/well/100 µL were added in a clear bottom 96-well plate. The absorbance was measured at 460 nm using a SpectraMax reader (Molecular Devices).
Cytotoxicity tests of HeLa cells in response to (A) SDS and (B) Staurosporine treatment were measured with Cell Meter™ Colorimetric WST-8 Cell Quantification Kit. HeLa cells at 10,000 cells/well/100 µL were seeded overnight in a black wall/clear bottom 96-well plate. Cells were treated with serially diluted SDS for 2 hours or Staurosporine for 4 hours. The absorbance was measured at 460 nm using a SpectraMax reader. 
Cell number was determined with Cell Meter™ Colorimetric WST-8 Cell Quantification Kit<strong>.</strong> Jurkat cells at 0 to 100,000 cells/well/100 µL were added in a clear bottom 96-well plate. The absorbance was measured at 460 nm using a SpectraMax reader (Molecular Devices).
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Catalog Number22770
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Telephone1-408-733-1055
Fax1-408-733-1304
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Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageRefrigerated (2-8 °C); Minimize light exposure
UNSPSC12352200
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OverviewpdfSDSpdfProtocol


Our Cell Meter™ assay kits are a set of tools for monitoring cell viability. There are a variety of parameters that can be used for monitoring cell viability. Cell Meter™ Colorimetric WST-8 Cell Quantification Kit uses water-soluble WST-8 tetrazolium salt to quantify the number of live cells. The water-soluble WST-8 tetrazolium salt produces a water-soluble orange formazan dye upon bioreduction in the presence of an electron carrier, 1-methoxy-5-methylphenazinium methyl sulfate. The kit is convenient and robust with a mix and read format. WST-8 solution is added directly to the test cells, no pre-mixing of components is required. WST-8 tetrazolium salt is reduced by cellular dehydrogenases to an orange formazan product that is soluble in tissue culture medium. The amount of formazan produced is directly proportional to the number of living cells by monitoring absorbance increase at 460 nm. The excellent stability and little cytotoxicity of WST-8 solution make the kit useful for the assays that require long incubation (such as 24 to 48 hours). Cell Meter™ Colorimetric WST-8 Cell Quantification Kit provides a sensitive colorimetric assay for the determination of the number of viable cells in the proliferation and cytotoxicity assays. The detection sensitivity is higher than any other tetrazolium salt-based assays such as MTT, XTT or MTS etc.

Platform


Absorbance microplate reader

Absorbance460 nm
Recommended plateClear bottom

Example protocol


AT A GLANCE

Protocol summary

  1. Prepare cells in a 96-well plate (100 µL/well)
  2. Add 10 µL of WST-8™ Solution to each well
  3. Incubate at 37°C for 1 - 4 hours
  4. Monitor absorbance at OD = 460 nm

Important notes
WST-8TM Solution is stable for more than one year if store at 4°C and protected from light. Store it at <-20°C for longer storage.

SAMPLE EXPERIMENTAL PROTOCOL

Cell Proliferation and Cytotoxicity Assay:

  1. Plate 5000 to 10,000 cells/well in a tissue culture microplate with clear bottom.

  2. 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 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.

  3. Add 10 µL/well (96-well plate) or 5 µL/well (384-well plate) of WST-8TM Solution to each well.

  4. 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.

  5. Monitor the absorbance increase with an absorbance plate reader at OD =460 nm.

Cell Counting Assay:

  1. Prepare cell culture in a tissue culture microplate with 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. 

  2. Add 10 µL/well (96-well plate) or 5 µL/well (384-well plate) of WST-8TM Solution to each well.

  3. 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.

  4. Monitor the absorbance increase with an absorbance plate reader at OD = 460 nm.

Citations


View all 2 citations: Citation Explorer
MicroRNA-30a suppresses self-renewal and tumorigenicity of glioma stem cells by blocking the NT5E-dependent Akt signaling pathway
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
miR-218 inhibits the proliferation of human glioma cells through downregulation of Yin Yang 1
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


View all 5 references: Citation Explorer
A sensitive WST-8-based bioassay for PEGylated granulocyte colony stimulating factor using the NFS-60 cell line
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
Comparison of the WST-8 colorimetric method and the CLSI broth microdilution method for susceptibility testing against drug-resistant bacteria
Authors: Tsukatani, T.; Suenaga, H.; Shiga, M.; Noguchi, K.; Ishiyama, M.; Ezoe, T.; Matsumoto, K.
Journal: J Microbiol Methods (2012): 160-6
WST-8 analysis of cell viability during osteogenesis of human mesenchymal stem cells
Authors: Stoddart, M. J.
Journal: Methods Mol Biol (2011): 21-5
monosodium salt (WST-8) rapid colorimetric assays for antimicrobial susceptibility testing of staphylococci and ESBL-producing clinical isolates
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
Reactivity of blood samples spotted onto filter papers in the WST-8 method for screening of G6PD deficiency
Authors: Arai, M.; Kosuge, K.; Kawamoto, F.; Matsuoka, H.
Journal: Acta Med Okayama (2006): 127-34