ApoLive-Glo™ Multiplex Assay Technical Manual

Literature # TM325

The ApoLive-Glo™ Multiplex Assay combines two assay chemistries to assess viability and caspase activation events within a single assay well. 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 (Figure 1). This live-cell protease becomes inactive upon loss of cell membrane integrity and leakage into the surrounding culture medium. The second part of the assay uses the Caspase-Glo™ Assay technology to detect caspase-3/7 activation, which is a key biomarker of apoptosis. The assay allows you to measure viability and apoptosis in the same sample well.