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Promega™ ApoLive-Glo™ Multiplex Assay DFS Item

The ApoLive-Glo™ Multiplex Assay measures both the number of viable cells as a marker of cytotoxicity and caspase activation as a marker of apoptosis within a single assay well to determine the mechanism of cell death.

Manufacturer:  Promega™ G6411

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Catalog No. PRG6411



Includes: GF-AFC Substrate, Assay Buffer, Caspase-Glo 3/7 Buffer, Caspase-Glo 3/7 Substrate

Description

Description

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

Specifications

Specifications

5 x 10mL
Fluorescence, Luminescence
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
Fluid
Determine mechanism of cell death, Profile compounds for cytotoxic risk
−30 to −10°C
Documents
Promotions

Promotions

U.S. Patents 7,416,854; 7,553,632 and other patents pending.
U.S. Patents 6,602,677 and 7,241,584, Australian Patents 754312 and 785294 and other patents and patents pending.
U.S. Patents 7,148,030 and 7,384,758, Australian Patent 2003216139 and other patents pending.
The method of recombinant expression of Coleoptera luciferase is covered by U.S. Patents 5,583,024, 5,674,713 and 5,700,673.