MultiTox-Glo Multiplex Cytotoxicity Assay

The MultiTox-Glo Multiplex Cytotoxicity Assay is a sequential-reagent-addition fluorescent and luminescent assay that measures the relative number of live and dead cells in cell populations. The MultiTox-Glo Assay sequentially measures two protease activities; one is a marker of viability, and the other is a marker of cytotoxicity. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant, peptide substrate (GF-AFC). This substrate enters intact cells, where it is cleaved by the live cell protease activity to release AFC and generate a fluorescent signa...

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The MultiTox-Glo Multiplex Cytotoxicity Assay is a sequential-reagent-addition fluorescent and luminescent assay that measures the relative number of live and dead cells in cell populations. The MultiTox-Glo Assay sequentially measures two protease activities; one is a marker of viability, and the other is a marker of cytotoxicity. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant, peptide substrate (GF-AFC). This substrate enters intact cells, where it is cleaved by the live cell protease activity to release AFC and generate a fluorescent signal that is proportional to the number of viable cells. The live-cell protease becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium. A second, luminogenic cell-impermeant peptide substrate (AAF-aminoluciferin) is used to measure dead-cell protease activity, which is released from cells that have lost membrane integrity. The liberated aminoluciferin product is measured as "glow type" luminescence generated by Ultra-Glo™ Recombinant Luciferase provided in the assay reagent.

The MultiTox-Glo Assay gives ratiometric, inversely correlated measures of cell viability and cytotoxicity, which correlate with established methods for measuring viability and cytotoxicity. The ratio of viable cells to dead cells is independent of cell number and, therefore, can be used to normalize data. Having complementary cell viability and cytotoxicity measures reduces errors associated with pipetting and cell clumping, as well as serving as an internal control to allow identification of errors resulting from chemical interference from test compounds or media components.

Features - Benefits

• Measure the Number of Live Cells and Dead Cells in Culture: Sequential-reagent-addition assay with a homogeneous "add-mix-measure" protocol.
• Normalize Data with a Built-In Internal Control: The ratio of the number of live cells/number of dead cells is independent of cell number and can be used to normalize data. This normalization makes results more comparable well-to-well, plate-to-plate and day-to-day.
• Immediately Identify More False-Positives and False-Negatives: Independent cell viability and cytotoxicity measurements serve as controls for each other. If test compounds interfere with one assay chemistry, the other serves as an internal control.
• Improve your Data: Reduce statistical probability of false-positives (or false-negatives), and eliminate fluorescence interference issues by luminescence readout.

Applications

• Determination of cell viability and cytotoxicity.
• Data normalization.

Notes

Using 100μl/assay in a 96-well plate format, Cat.# G9270 is sufficient to perform 100 assays; Cat.# G9271, 500 assays; Cat.# G9272, 1,000 assays. Using 25μl per well in a 384-well plate format, Cat.# G9270 is sufficient to perform 400 assays; Cat.# G9271, 2,000 assays; Cat.# G9272, 4,000 assays.

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