Science Catalog 2012 Life Cell Signaling ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay Product ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay For Research Use Only. Not for Use in Diagnostic Procedures. Description: The ONE-Glo™ + Tox Assay combines luciferase assay chemistry with a cell viability marker to better understand reporter gene expression in the context of cell health. The assay uses a two-step, addition-only process to make these measurements in a single well of a plate, negating the need to run parallel assays. The first part of the assay is a nonlytic fluorescence assay (CellTiter-Fluor™ Cell Viability Assay) that measures the relative number of live cells in a culture population after experimental manipulation. The CellTiter-Fluor™ Assay measures a conserved and constitutive protease activity within live cells and therefore serves as a marker of cell viability. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cellpermeant peptide substrate (glycylphenylalanyl-aminofluorocoumarin; GF-AFC). The substrate enters intact cells where it is cleaved by the live-cell protease to generate a fluorescent signal proportional to the number of living cells. This live-cell protease becomes inactive upon loss of cell membrane integrity and leakage into the surrounding culture medium. Fluorescence of the free AFC fluorophore is measured with a microplate reader or CCD imager using an excitation wavelength of 380–400nm and emission wavelength of 505nm. The second part of the assay uses the ONE-Glo™ Luciferase Assay System to quantify firefly luciferase reporter gene expression from cells made to express this reporter enzyme. The ONE-Glo™ Luciferase Assay Buffer and ONE-Glo™ Luciferase Assay Substrate, provided with this system, are combined to form the ONE-Glo™ Reagent. Ideally suited for high- and ultrahigh-throughput applications, the ONE-Glo™ Assay contains a new fluoroluciferin substrate, resulting in a reagent that is more stable, more tolerant to sample components, and has less odor than standard luciferase assay reagents. Luminescence is measured with a microplate reader or CCD imager. CF3 GF–N H O O LIVE-CELL SUBSTRATE: cell-permeant fluorogenic substrate for the live-cell protease (Gly-Phe-AFCoumarin) Live-cell protease substrate can cross the cell membrane. Size Cat.# Price (Fr) 1 plate E7110 Pls. Enq. 10 plates E7120 Pls. Enq. Features: • Efficient: Measure cell viability and firefly luciferase gene expression within the same assay well. Acquire more data by performing two measurements on cells in a single assay well. • Better Biology: Understand reporter gene expression in the context of cell viability. • Easy to Perform: The assay uses a simple sequential “add-mix-read” format. • Flexible and Automation-Friendly: The volumes of each assay component can be scaled to meet throughput needs and is amenable to automation up to 1,536-well format. Storage Conditions: Store the ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay components at –20°C. Please refer to the Technical Manual for other storage options. Protocol ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay Technical Manual Steady-Glo® Luciferase Assay System Product Steady-Glo® Luciferase Assay System For Research Use Only. Not for Use in Diagnostic Procedures. Description: High-throughput quantitation of firefly (Photinus pyralis) luciferase expression in mammalian cells is commonly performed by batch processing of 96- and 384-well plates. Steady-Glo® Luciferase Assay System is designed for this purpose by providing long-lived luminescence when added to cultured cells. The homogeneous assay provides signal half-lives of over 5 hours in commonly used cell culture media without prior sample processing. Throughput rates of several thousand samples per hour may be achieved with high reproducibility under standard laboratory conditions. Features: • Greater Light Output: Results in greater assay sensitivity than other leading extended-lifetime firefly luciferase assay reagents. • Improved Assay Precision: Less sensitive to mixing conditions for improved assay reproducibility in multiwell plates. This is particularly useful in 384-well plates. • Convenient: Simply mix buffer with lyophilized substrate and use; there is no need for thawing or measuring before use. • Quick: No need to remove culture medium or wash cells prior to adding assay reagent. Cells may be grown and assayed directly within the same multiwell plate. • Easy to Use: Simply add reagent, which contains a cell lysis component, wait 5 minutes and measure luminescence. Active Live-Cell Protease H2N Nucleus Live Cell The CellTiter-Fluor™ Cell Viability Assay chemistry portion of the One-Glo™ + Tox Assay is a nonlytic fluorescent assay that measures the relative number of live cells in a culture population after experimental manipulation. 264 For complete and up-to-date product information visit: www.promega.com/catalog GF CF3 O O • Robust: Compatible with many tissue culture media, including those containing up to 10% serum. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store Steady-Glo® Luciferase Assay Substrate at –20°C. Store Steady-Glo® Luciferase Assay Buffer below 25°C. Protocol Steady-Glo® Luciferase Assay System Technical Manual Part# TM051 Size Cat.# Price (Fr) 10 ml E2510 172.00 100 ml E2520 1204.00 10 × 100 ml E2550 Pls. Enq. Part# TM356 6995MB Pagina 267
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