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QIA127 | Rapid Cell Proliferation Kit

QIA127
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      Descripción

      Replacement Information

      Tabla espec. clave

      Detection Methods
      Colorimetric

      Precios y disponibilidad

      Número de referenciaDisponiblidad Embalaje Cant./Env. Precio Cantidad
      US1QIA127-1KIT
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      Disponibilidad a confirmarDisponibilidad a confirmar
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          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.
          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

          Documentation

          Ficha datos de seguridad (MSDS)

          Título

          Ficha técnica de seguridad del material (MSDS) 

          Certificados de análisis

          CargoNúmero de lote
          QIA127

          Referencias bibliográficas

          Visión general referencias
          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.
          Protocolo de usuario

          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.