Science Catalog 2012 Life Cell Signaling Fluorescent Cell Viability Assay Product CellTiter-Fluor™ Cell Viability Assay For Laboratory Use. Description: The CellTiter-Fluor™ Cell Viability Assay is a nonlytic, singlereagent-addition fluorescence assay that measures the relative number of viable cells in a population. The assay is based on measurement of a conserved and constitutive protease activity within live cells and therefore serves as a biomarker of cell viability. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant, peptide substrate (Gly-Phe-AFC). The substrate enters intact cells, where it is cleaved by the live-cell protease activity to generate a fluorescent signal proportional to the number of living cells. The live-cell protease becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. The CellTiter-Fluor™ Assay also can be used in a single-well, sequential, multiplex format with other downstream assay chemistries to normalize data by cell number. Data from the assay can serve as an internal control and allow identification of errors resulting from cell clumping or compound cytotoxicity. The assay is compatible with many Promega luminescence assays or spectrally distinct fluorescence assay methods, such as measuring caspase activation, reporter gene expression or orthogonal measures of viability. Features: • Obtain Better Data from Every Well: The assay can be performed in multiplex with many Promega luminescence assays or spectrally distinct fluorescence assays. • Normalize Data for Cell Number: Normalizing data for live-cell number makes results more comparable well-to-well, plate-to-plate, day-to-day. • Save on Cell Culture Costs: Multiplexing assays in the same well eliminates parallel plate processing, thus reducing cell culture costs. Storage Conditions: Store at –20°C. Protocol CellTiter-Fluor™ Cell Viability Assay Technical Bulletin CellTiter 96® AQueous One Solution Cell Proliferation Assay (MTS) Product CellTiter 96® AQueous One Solution Cell Proliferation Assay For Laboratory Use. Description: The CellTiter 96® AQueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation, cytotoxicity or chemosensitivity assays. The CellTiter 96® AQueous One Solution Reagent contains a tetrazolium compound [3-(4,5-dimethylthiazol2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and an electron coupling reagent (phenazine ethosulfate; PES). PES has enhanced chemical stability, which allows it to be combined with MTS to form a stable solution. The CellTiter 96® AQueous Assay uses phenazine methosulfate (PMS) as the electron coupling reagent, and PMS Solution and MTS Solution are supplied separately. PES has enhanced chemical stability, which allows it to be combined with MTS to form a stable solution. Assays are performed by adding a small amount of the CellTiter 96® AQueous One Solution Reagent directly to culture wells, incubating for 1–4 hours and then recording absorbance at 490nm with a 96-well plate reader. The quantity of formazan product as measured by the amount of 490nm absorbance is directly proportional to the number of living cells in culture. Size Cat.# Price (Fr) 200 assays G3582 168.00 1,000 assays G3580 504.00 5,000 assays G3581 1512.00 0.50 0.00 Part# TB371 Comparison of MTS and [3H]thymidine Assays Proliferation of HT-2 Cells Stimulated with GM-CSF 2.00 1.50 1.00 MTS [3H]-Thy 4 0 .01 .04 .16 ng/ml GM-CSF the CellTiter 96® AQueous Cell Proliferation Assay and a [3H]thymidine incorporation assay. Similar results were obtained with both assays. Measurement of GM-CSF-stimulated proliferation in HT-2 cells using .62 2.5 10 0 16 12 8 Size Cat.# Price (Fr) 10 ml G6080 175.00 5 × 10 ml G6081 613.00 2 × 50 ml G6082 1043.00 If you currently use a [3H]-thymidine incorporation assay, addition of the CellTiter 96® AQueous One Solution Reagent can be substituted for the pulse of [3H]-thymidine at the time point in the assay when the pulse of radioactive thymidine is usually added. Previous bioassay data comparing [3H]-thymidine incorporation to the MTS-based CellTiter 96® AQueous Assay and the original MTT-based CellTiter 96® Assay demonstrate that tetrazolium reagents can be substituted for [3H]-thymidine incorporation. Features: • Simplify Colorimetric Viability Assays: “Add-incubate-measure” format enables design of homogeneous high-throughput screening assays. • Use a Single Solution: Use as a single solution, filter sterilized and ready to add to assay plates (unlike MTT). • Perform Fewer Steps: Perform the assay in 96-well plates with no washing or cell harvesting. Also eliminates solubilization steps normally required for MTT assays. • Gain Flexibility: Plates can be read and returned to incubator for further color development (unlike MTT). • Avoid Organic Solvents: Requires no volatile organic solvent to solubilize the formazan product (unlike MTT). • Non-Radioactive: Requires no scintillation cocktail or radioactive waste disposal (unlike [3H]-thymidine incorporation assays). • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –20°C, protected from light. Protocol CellTiter 96® AQueous One Solution Cell Proliferation Assay System Technical Bulletin Part# TB245 36 For complete and up-to-date product information visit: www.promega.com/catalog Corrected Ab. 490nm 2372MA08_8A CPM (x 10-3) Pagina 39
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