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Products >  Cell_Biology >  Cell_Proliferation_and_Viability >  EIA_Kits >  Premix_WST-1_Cell_Proliferation_Assay_System

Premix WST-1 Cell Proliferation Assay System

The Premix WST-1 Cell Proliferation Kit allows fast and easy colorimetric measurement of cell proliferation and viability of cells cultured in 96-well plates. This assay is based on the cleavage of tetrazolium salts (WST-1) to soluble formazan dye by mitochondrial succinate-tetrazolium reductase in viable cells. As a cell population increases, an increase in the amount of reductase present in the culture supernatant results in a concomitant increase in conversion of WST-1 to formazan dye. Thus, the quantity of formazan dye is directly proportional to the number of metabolically active cells in the medium. Formazan dye formed by metabolically active cells can be quantified by measuring its absorbance (440 nm) using a microtiter plate reader.

This kit provides a non-radioactive and spectrophotometric method of determining cell proliferation and viability without harvesting or washing steps, and can be performed in a single multi-well plate. The Premix WST-1 is supplied as a ready-to-use solution containing WST-1 and an electron coupling reagent, diluted in sterile phosphate buffered saline. Premix WST-1 is diluted 1:10 with 100-200 µL cultured cells/well. The kit supplies sufficient reagents for 2,500 assays.
At-A-Glance Documents Images & Data

Features

  • Safe to use: No radioactive isotopes are required; no volatile organic solvent is required for solubilization
  • Accuracy: Absorbance strongly correlates to the number of viable cells
  • Sensitivity: More sensitive than using MTT
  • Ease of Use: Supplied as a ready-to-use, sterile solution
    -- Short reaction time
    -- The entire assay is performed in one microtiter plate
    -- No need for washing, harvesting or farmazan dye solubilization
    -- Processing a large number of samples is possible using a multiwell-ELISA reader
    -- No isotope disposal and radiation safety paperwork
  • Flexibility: Plates can be read and returned several times to the incubator for further color development

Applications

  • Measurement of cell proliferation due to growth factors, cytokines, mitogens, and nutrients
  • Analysis of cytotoxic and cytostatic compounds
  • Evaluation of factors which inhibit cell growth

Components

PreMix WST-1*1 bottle x 25 mL

* The PreMix WST-1 is a ready-to-use solution containing
WST-1 and an electron coupling reagent, diluted in phosphate
buffered saline, sterile.

Storage

Shipped at –20°C. Stored at –20°C, protected from light.

If a precipitate is observed when dissolving this reagent, redissolve any precipitate by warming at 37°C for several minutes with gentle shaking. Once thawed, the solution can be stored at 4°C and protected from light for several weeks. For longer storage, Takara recommends storing in aliquots at –20°C. Do not repeat freeze-thaw cycle.

Principle

Tetrasolium salt (WST-1) is cleaved to soluble formazan dye by the succinate-tetrazolium reductase (EC 1.3.99.1), which exists in mitrochondrial respiratory chain and is active only in viabale cells. Total activity of this mitochondrial dehydrogenase in a sample rises with the increase of viable cells. As the increase of enzyme activity leads to an increase of the production of formazan dye, the quantity of formazan dye is related directly with the number of metabolically active cells in the medium. The formazan dye formed by metabolically active cell can be quantitated by measuring its absorbance by ELISA reader. The absorbance of formazan dye solution is in direct proportion to the number of viable cells. This product is also designed for non-radioactive and spectrophotometric quantification of cell growth and viability in proliferation and chemosensitivity assay.

References

  1. Cook, J. A. and Mitchell, J.B. (1989) Anal. Biochem 179, 1-7.
  2. Mosmann, T. (1983) J. Immunol. Methods 65, 55-63.
  3. Carmichael, J. et. al. (1987) Cancer Res. 47, 936-942.
  4. Vistica, D. T. et. al. (1991) Cancer Res. 51, 2515-2520.
  5. Scudiero, D. A. et. al. (1988) Cancer Res. 48, 4827-4833.
  6. Weislow, O.S. et al. (1989) J. Natl. Cancer Inst. 81, 577-586.
  7. Roehm, N. W. et al. (1991) J. Immunol. Methods 142, 257-265.
  8. Cory, A. H. et al. (1991) Cancer Commun. 3, 207-212.



 
 
Products
Cat. # Product Package Size Price
MK400 Premix WST-1 Cell Proliferation Assay System 2,500 Tests $399.00
 

 


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