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CELLPRO-RO Roche

Cell Proliferation Reagent WST-1

Synonym: cell proliferation reagent, wst-1

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Properties

Quality Level   100
form   solution
usage   sufficient for ≤2,500 tests (11644807001)
  sufficient for ≤800 tests (05015944001)
packaging   bottle of 25 mL (11644807001)
  bottle of 8 mL (05015944001)
mfr. no.   Roche
storage condition   protect from light
application(s)   cell analysis: suitable
  detection: suitable
  protein quantification: suitable
optimum pH   8.0(for physiological conditions our product is buffered pH 7.3)
λmax   440-480 nm
detection method   colorimetric
shipped in   dry ice
storage temp.   −20°C

Description

General description

Contents
Ready-to-use solution, containing WST-1 and an electron coupling reagent.

Cellular proliferation requires the replication of genomic DNA. Thus, monitoring DNA synthesis is an indirect parameter of cell proliferation, as well as being suitable for the study of the regulation of DNA synthesis. [3H]-thymidine has traditionally been used to label the DNA of replicating (cycling) cells. To circumvent the disadvantages associated with the use of the [3H]-thymidine, nonradioactive alternatives based on 5-bromo-2′-deoxyuridine (BrdU) have been developed. The use of chemiluminescence technology provides enhanced sensitivity and a broad measurement range.

Colorimetric assay (WST-1 based) for the nonradioactive quantification of cell proliferation, cell viability, and cytotoxicity.

Application

The Cell Proliferation Reagent WST-1 is used for the nonradioactive, spectrophotometric quantification of cell proliferation and viability in cell populations using the 96-well-plate format. Measurement of cell proliferation in response to growth factors, cytokines, mitogens, and nutrients.
• Analysis of cytotoxic and cytostatic compounds, such as anti-cancer drugs and other pharmaceutical compounds
• Assessment of growth inhibitory antibodies and physiological mediators

Features and Benefits

• Convenient: Benefit from a ready-to-use reagent.
• Safe and Easy: Eliminate radioactive isotopes, washing steps, and additional reagents.
• Accurate: The absorbance obtained strongly correlates to the cell number.
• Sensitive: Detect low cell numbers.
• Fast: Process a large number of samples using a multi-well ELISA reader.

Principle

The stable tetrazolium salt WST-1 is cleaved to a soluble formazan by a complex cellular mechanism that occurs primarily at the cell surface. This bioreduction is largely dependent on the glycolytic production of NAD(P)H in viable cells. Therefore, the amount of formazan dye formed directly correlates to the number of metabolically active cells in the culture.
Cells grown in a 96-well tissue culture plate are incubated with the WST-1 reagent for 0.5 - 4 hours. After this incubation period, the formazan dye formed is quantitated with a scanning multi-well spectrophotometer (ELISA reader). The measured absorbance directly correlates to the number of viable cells.
Cell proliferation and viability assays are of particular importance for routine applications in cell biology. Tetrazolium salts (e.g., MTT, XTT, WST-1) are particularly useful for this type of analysis. Tetrazolium salts are cleaved to formazan by the succinate-tetrazolium reductase system (EC 1.3.99.1) which belongs to the respiratory chain of the mitochondria, and is only active in metabolically intact cells.

Preparation Note

Working concentration: For best results, add 10 μl/well Cell Proliferation Reagent WST-1 to the cells already cultured in 100 μl/well (1:10 final dilution).
Using the 100 μl/well cell culture volume, one vial will be sufficient to perform 2500 tests (25 microplates).
Note: If the cells are cultured in 200 μl/well, add 20 μl/well Cell Proliferation Reagent WST-1.
Storage conditions (working solution): 2 to 8 °C
Note: When precipitates or turbidity are observed upon thawing, warm up the solution to 37 °C for 2 to 10 minutes and agitate to dissolve the precipitates.Centrifugation is not recommended because the working concentration would decrease. After being dissolved, the WST-1 reagent can be used without any limitations. Please store as follows:
Once thawed, store at 2 to 8 °C, protected from light, for up to four weeks. However please note that the solution may become viscous. If so, warm up the solution to 37 °C for 2 to 10 minutes as described above.

Long storage
For long storage, aliquots of WST-1 can be stored in plastic tubes at -15 to -25 °C until the expiry date.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Cell Viability and Proliferation Assays

Assays to measure cellular proliferation, cell viability, and cytotoxicity are commonly used to monitor the response and health of cells in culture after treatment with various stimuli. The proper ch...
Keywords: Cancer, Cell culture, Cell disruption, Cell proliferation, Click chemistry, Enzyme-linked immunosorbent assay, Flow cytometry, Glycolysis, Immunocytochemistry, Immunohistochemistry, Indicators, Microscopy, Nucleic acid denaturation, Reductions, Solvents, Western blot

Protocols

Protocol Guide: WST-1 Assay for Cell Proliferation and Viability

The measurement of cell proliferation and cell viability has become a key technique in the life sciences. The need for sensitive, reliable, fast and easy methods has led to the development of several...
Keywords: Absorption, Antibiotics, Apoptosis, Cancer, Cell culture, Cell proliferation, Enzyme-linked immunosorbent assay, Growth factors, Pharmaceutical, Phase transitions

Peer-Reviewed Papers
15

References

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