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MultiTox-Glo Multiplex Cytotoxicity Assay

by Promega
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  • A Bioluminescent and Fluorescent Detection 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 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. A luminescent readout allows for a reduction in the statistical probability of false-positives (or false-negatives) and eliminates fluorescence interference issues.

    The Biology of the MultiTox-Glo Multiplex Cytotoxicity Assay


    The MultiTox-Glo Assay Measures Distinct and Differential Protease Activities

    Viability and Cytotoxicity Measurements Are Inversely Correlated and Ratiometric

    Notes

    Using 100ul/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 25ul 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.
  • Protocols

    Complete Protocol

    Download PDF

    MultiTox-Glo Multiplex Cytotoxicity Assay Technical Bulletin

    PDF (471 KB)

  • Certificate of Analysis

    Lookup Certificate of Analysis

    Storage Conditions

    -30C TO -10C

    Use Restrictions

    For Research Use Only. Not for Use in Diagnostic Procedures.

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