MultiTox-Fluor Multiplex Cytotoxicity Assay

The MultiTox-Fluor Multiplex Cytotoxicity Assay is a single-reagent-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 activ...

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The MultiTox-Fluor Multiplex Cytotoxicity Assay is a single-reagent-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 - Benefits

• Measure the Number of Live and Dead Cells in Culture: Homogeneous, "add-mix-measure" 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. Data normalization for cell number makes results more comparable well-to-well, plate-to-plate, day-to-day.
• Reduce False-Positive and -Negative Results: Complementary live- and 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.

Applications

• Determine viability, cytotoxicity or both in the same well.
• Multiplex with Promega luminescent cell-based assays.

Notes

G9200 contains sufficient reagents for 100 assays at 100μl/assay in a 96-well format or 400 assays at 25μl/assay in a 384-well format. G9201 contains sufficient reagents for 500 assays at 100μl/assay in a 96-well format or 2,000 assays at 25μl/assay in a 384-well format. G9202 contains sufficient reagents for 1,000 assays at 100μl/assay in a 96-well format or 4,000 assays at 25μl/assay in a 384-well format.

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