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QIA127 Rapid Cell Proliferation Kit

Rapid Cell Proliferation Kit MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
QIA127
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Přehled

Replacement Information

Tabulka spec. kláve

Detection Methods
Colorimetric

Products

Katalogové čísloBalení ks/bal.
QIA127-1KIT Skleněná láhev 1 kit
Description
OverviewAssay measures the increased activity of cellular mitochondrial dehydrogenases that can cleave the tetrazolium dye WST-1 to formazan. The formazan formation is then quantified by measuring the change in absorbance at 450 nm in a microplate reader. The activity of mitochondrial dehydrogenases is proportional to cell number. No washing, harvesting, or solubilization steps are required.
Catalogue NumberQIA127
Brand Family Calbiochem®
Materials Required but Not Delivered• Pipettors with disposable tips
• Tissue culture grade, flat-bottom 96-well plate
• Spectrophotometer capable of measuring absorbance in 96-well plates at a wavelength of 440-460 nm
References
ReferencesIshiyama, M., et al. 1995. In Vitro Toxicology 8, 187.
Liu, S.O., et al 1995. Nat. Med. 1, 267.
Ishiyama, M., et al. 1993. Chem. Pharm. Bull. 41, 1118.
Product Information
Detection methodColorimetric
Form500 Tests
Format96-well plate
Kit containsWST-1, 1-Methoxy PMS, and a user protocol.
Quality LevelMQ100
Applications
Biological Information
Assay range1000 - 50,000 Cells
Assay time0.5 - 4 h
Sample TypeCultured cells
Physicochemical Information
Sensitivity1000 cells (adherent); 2500 cells (nonadherent)
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Intended useThe Calbiochem® Rapid Cell Proliferation Kit is designed to determine the number of viable cells in proliferation, or cytotoxicity studies, based on the cleavage of the tetrazolium salt WST-1 by mitochondrial dehydrogenases in viable cells. The assay can be used to measure cell proliferation in response to growth factors or cytokines, or to assess cytotoxic compounds.
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Multiple Toxicity Values, refer to MSDS
Storage -20°C
Storage ConditionsUpon arrival store the entire contents of the kit at -20°C.
Protect from Light Protect from light
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Kit containsWST-1, 1-Methoxy PMS, and a user protocol.
Specifications
Global Trade Item Number
Katalogové číslo GTIN
QIA127-1KIT 07790788054021

Documentation

Rapid Cell Proliferation Kit MSDS

Title

Safety Data Sheet (SDS) 

Rapid Cell Proliferation Kit Certificates of Analysis

TitleLot Number
QIA127

References

Přehled odkazů
Ishiyama, M., et al. 1995. In Vitro Toxicology 8, 187.
Liu, S.O., et al 1995. Nat. Med. 1, 267.
Ishiyama, M., et al. 1993. Chem. Pharm. Bull. 41, 1118.
User Protocol

Revision11-September-2008 RJH
Form500 Tests
Format96-well plate
Detection methodColorimetric
StorageUpon arrival store the entire contents of the kit at -20°C.
Intended useThe Calbiochem® Rapid Cell Proliferation Kit is designed to determine the number of viable cells in proliferation, or cytotoxicity studies, based on the cleavage of the tetrazolium salt WST-1 by mitochondrial dehydrogenases in viable cells. The assay can be used to measure cell proliferation in response to growth factors or cytokines, or to assess cytotoxic compounds.
BackgroundCell proliferation assays are widely used in cell biology to study growth factors, cytokines, and media components, to screen cytotoxic agents, and for lymphocyte activation. The use of tetrazolium salts such as MTT commenced in the 1950s, and is based on the fact that live cells reduce tetrazolium salts into colored formazan compounds. The biochemical procedure is based on the activity of mitochondrial enzymes, which are inactivated shortly after cell death. The new cell proliferation reagent WST-1 has several advantages as compared to MTT. WST-1 yields water-soluble cleavage products like XTT, which can be measured without an additional solubilization step, and WST-1 has a wider range than MTT or XTT.
Principles of the assayThe Calbiochem® Rapid Cell Proliferation Kit provides a colorimetric assay for the fast and convenient determination of viable cell numbers in cell proliferation and cell cytotoxicity assays. After mixing the two components supplied in the kit the mixed solution is added to the wells of a 96-well plate (tissue culture plate) that contains cultivated cells. The intensity of the dye is proportional to the number of viable cells and the absorbance is read with a microplate reader at 440-460 nm after a 0.5-4 h incubation at 37°C.
Materials provided• WST-1 Reagent (Kit Component No. JA7700-1EA): 1 vial, supplied as a powder in an amber glass bottle
• Electron Mediator Solution (EMS) (Kit Component No. JA7701-5ML): 1 vial, 5 ml in an amber glass bottle
Materials Required but not provided• Pipettors with disposable tips
• Tissue culture grade, flat-bottom 96-well plate
• Spectrophotometer capable of measuring absorbance in 96-well plates at a wavelength of 440-460 nm
Reagent preparation• WST-1 Labeling Mixture: Thaw the EMS solution at room temperature or in a 37°C water bath. Add the entire contents of this solution to the bottle containing the WST-1 reagent. The mixed solution can be stored at 4°C for several weeks. For long-term storage (several months) aliquot and store at -20°C. Protect from light.
Detailed protocol1. Grow cells in a tissue culture grade, flat-bottom plate in 100 µl of culture medium per well in a CO2 tissue culture incubator. For cell stimulation assays a starting concentration of 5,000-25,000 cells per well is recommended, for cytotoxicity assays the recommended number of cells per well is 25,000-150,000.
2. After incubation add 10 µl of the WST-1 labeling mixture, prepared as above, to each well. Mix briefly by shaking gently for several min.
3. Incubate adherent cells for 1-2 h and suspension cells for 3-4 h.
4. Measure the absorbance of the samples using a microplate reader at a wavelength of 440-460 nm. If a reference wavelength is to be subtracted, a filter above 600 nm is recommended. The background absorbance is dependent upon the culture medium, pH, incubation time, and time of exposure to light. The typical background absorbance after a 2 h incubation is 0.1-0.2 absorbance units.
Example data

Figure 1: Example

NIH3T3 cells incubated with WST-1 labeling mixture for 2.5 h at 37°C as described above.

Figure 2: No. NIH3T3 Cells vs. Abs 450 nm

NIH3T3 cells incubated with WST-1 labeling mixture for 2.5 h at 37°C as described previously.

Figure 3: No. HT-1080 Cells vs. Abs 450 nm

HT-1080 cells incubated for 20 h followed by the addition of WST-1 labeling mixture. After additional 1 (Δ), 2.5 (⊄), and 4 h (•) incubations the absorbance was read at 450 nm.

Figure 4: Inductions of Apoptosis in Jurkat Cells

100,000 Jurkat cells per well were incubated overnight at 37°C in the presence of 0, 1, or 5 µg/ml camptothecin in RPMI medium containing 2% FBS. After the incubation 10 µl of the WST-1 labeling mixture was added and absorbance was read after a 2.5 h incubation at 37°C.
Sensitivity1000 cells (adherent); 2500 cells (nonadherent)
Assay Range1000 - 50,000 Cells
Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
Interactive Pathways™ is a trademark of EMD Chemicals, Inc.