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Promega™ ApoLive-Glo™ Multiplex Assay


Measures both the number of viable cells as a marker of cytotoxicity and caspase activation as a marker of apoptosis in a single well to determine the mechanism of cell death

Manufacturer: promega™  G6411

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Catalog No. PRG6411

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Description & Specifications

Specifications

Quantity 5 x 10mL
Format Fluid
Detection Method Fluorescence, Luminescence
For Use With (Application) Determine mechanism of cell death, Profile compounds for cytotoxic risk
Kit Contents 1 x 50μL GF-AFC substrate, 2 x 10mL Assay Buffer, 5 x 10mL Caspase-Glo™ 3/7 Buffer, 5 x 1 bottle Caspase-Glo™ 3/7 substrate
Storage Requirements -20°C

The first part of the assay measures the activity of a protease marker of cell viability. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant, peptide substrate (glycyl-phenylalanyl-amino fluorocoumarin; GF-AFC). The substrate enters intact cells, where it is cleaved by the live-cell protease activity to generate a fluorescent signal proportional to the number of living cells. The second part of the assay uses the Caspase-Glo™ Assay technology to detect caspase activation, a key biomarker of apoptosis. Adding the Caspase-Glo 3/7 Reagent in an “add-mix-measure” format results in cell lysis, followed by caspase cleavage of the substrate and generation of a “glow-type” luminescent signal that is proportional to the amount of caspase activity present.

  • Accurately determines the mechanism of cell death in less time with less sample
  • Easy-to-use “add-mix-measure” format
  • Permits the normalization of caspase data with viability control, which is useful for determining the extent of caspase acivation and for normalizing cell numbers
  • Scalable and easily automated in 96- and 384-well plates
  • Reveals cell death even if the window of caspase activity is missed
  • Allows further multiplexing with other assays with spectrally distinct fluorescent assay chemistries due to the nonlytic nature of the first step of the assay