Cell Signaling MultiTox-Fluor Multiplex Cytotoxicity Assay Product MultiTox-Fluor Multiplex Cytotoxicity Assay For Laboratory Use. Description: The MultiTox-Fluor Multiplex Cytotoxicity Assay is a singlereagent-addition, homogeneous, fluorescent assay that measures the number of live and dead cells simultaneously in culture wells. The assay simultaneously measures cell viability and cytotoxicity by detecting two distinct protease activities. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant peptide substrate (GF-AFC Substrate). The substrate enters intact cells where it is cleaved to generate a fluorescent signal proportional to the number of living cells. This live-cell protease activity marker becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. A second, cell-impermeant, fluorogenic peptide substrate (bis-AAF-R110 Substrate) is used to measure dead-cell protease activity that has been released from cells that have lost membrane integrity. Features: • Measure the Number of Live and Dead Cells in Culture: “Add-mixmeasure” protocol eliminates parallel plate processing and reduces cell culture costs. • Normalize Data for Cell Number: The ratio of live:dead cells is independent of cell number and normalizes data, making results more comparable well-to-well, plate-to-plate, day-to-day. • Reduce False-Positive and -Negative Results: Complementary liveand dead-cell measures with independent chemistries serve as internal controls for each other. • Get More Data from Every Well: Multiplex the MultiTox-Fluor Assay with most Promega bioluminescent cell-based apoptosis or genetic reporter assays. • Reduce Assay Variability: The homogeneous “add-mix-measure” protocol avoids the cumulative error associated with multistep protocols. Storage Conditions: Store at –20°C. Protocol MultiTox-Fluor Multiplex Cytotoxicity Assay Part# TB348 2) Add substrates.tes. GF– LIVE-CELL SUBSTRA cell permeant fluorogenic substrate for the live-cellf protease (Gly-Phe-AFCoumarin) ELL SUBSTRASTRATE: meant fluorogenic h li ll AAF AAF– cell impermeant substrate h dd ll DEAD-CELL SUBSTRA cell impermeant substrate f for the dead-cell protease (Ala-Ala-Phe-rhodamine 110 or Ala-Ala-Phe-Aminoluciferin) Active Live-Cell Protease Nucleus Live Cell GF+ Live-cell protease substrate can cross the cell membrane; dead cell protease substrate cannot. Active Dead-Cell Protease Active Dead-Cell Protease AAF+ Dead Cell Inactive Live-Cell Protease 3) Measure fluorescence/luminescence. 100% Live Cells 50% Live / 50% Dead Cells 100% Dead Cells 0% Live Cell Nucleus Live Cell Nucleus % Live / 50% D Live Cell Nucleus Deead Cells Dead Cell Dead Cell 100% Dead Cells Dead Cell Dead Cell Size Cat.# Price (Fr) 10 ml G9200 340.00 5 × 10 ml G9201 1190.00 2 × 50 ml G9202 2023.00 MultiTox Assay Protocol Live Cell Active Live-Cell Protease Active Dead-Cell Protease Nucleus 1) Treat cells with the potential cytotoxic agent. with the totoxic c agent. e Live Cell Active Live-Cell Protease Active Dead-Cell Protease Nucleus Active Dead-Cell Protease Dead Cell Inactive Live-Cell Live Cell Active Live-Cell Protease Active Dead-Cell Protease Nucleus 2 2 2 2 1 1 1 0 Live Cells Dead Cells 0 Live Cells Dead Cells 0 Live Cells Dead Cells Fluorescence at 520nm (rhodamine 110-red) or 505nm (coumarin-blue) Overview of the MultiTox-Fluor Multiplex Cytotoxicity Assay protocol. For complete and up-to-date product information visit: www.promega.com/catalog 39 Cell Health Assays 6581MB Pagina 42
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