Invitrogen™ CyQUANT™ XTT Cell Viability Assay

Description

• Kit consists of two reagents, XTT Reagent and Electron Coupling Reagent
• XTT Reagent is used to assess cell viability as a function of cellular redox potential, and the electron coupling reagent improves the dynamic range of the assay
• The assay is sufficient for ten 96-well plates and consists of 10 bottles of XTT Reagent and 10 tubes of Electron Coupling Reagent
• One bottle of each (sufficient for one 96 well plate) are thawed, mixed together, added to cells, and incubated
• The viability signal is detected using an absorbance-based microplate reader
• In actively respiring viable mammalian cells, the water-soluble XTT reagent is reduced and converted to an orange-colored formazan product
• Features of the CyQUANT™ XTT Cell Viability Assay include: • Highly reproducible, absorbance-based viability assay • Improved dynamic range and sensitivity compared to other colorimetric viability assays • Simple mix-and-read format, configured for one 96-well plate at a time • Non-toxic, continuous assay—multiple time points can be collected, no cell lysis Measuring changes in cell viability is a fundamental method for assessing cell health, determining genotoxicity, and evaluating anti-cancer drugs
• In addition to being cost effective, colorimetric viability assays generate consistent results with low well-to-well variability
• Unlike other commercially available colorimetric mammalian cell viability assays, XTT-based assays display a high dynamic range and low variability
• In addition to improved performance, the assay is a continuous assay and requires no cell lysis or solubilization
• The CyQUANT™ XTT Cell Viability Assay is a complete and optimized kit for the detection of mammalian cell viability
• The assay kit includes XTT Reagent (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) and Electron Coupling Reagent
• The XTT reagent, which is a tetrazolium-based compound, is sensitive to cellular redox potential and in the presence of actively respiring cells converts from a water-soluble compound to an orange-colored formazan product
• The sensitivity and consistency of the assay is significantly increased when used with the electron coupling reagent
• Since no cell lysis or solubilization are required for detection, the XTT assay can be employed as a continuous assay and several time points can be analyzed
• Additionally, the CyQUANT™ XTT assay generates viability results from various types of cell lines, including primary and suspension cells
• For each 96-well plate, one bottle of XTT Reagent and one tube of Electron Coupling Reagent are thawed, mixed to create a stock solution, and added to cells, with cell viability detected after a 2–4 hour incubation
• The assay kit supplies all the materials needed for ten 96-well plates
• It is recommended that the XTT/Electron Coupling reagent stock solution be used soon after mixing
• Assay performance is compromised and sensitivity is reduced if the stock solution is kept at room temperature for extended periods of time or subject to cycles of freeze/thaw
• To ensure assay performance and reproducibility, the CyQUANT™ XTT Cell Viability Assay consists of separate XTT and Electron Coupling reagents that are mixed.