ApoTox-Glo Triplex Assay

Description

• Overview
• Protocols
• Specifications
• Resources

• Three Effective Assays in One Single Product

  The ApoTox-Glo™ Triplex Assay combines three assay chemistries to easily assess viability, cytotoxicity and apoptosis events in the same cell-based assay well. First, viability and cytotoxicity are determined by measuring two differential protease biomarkers simultaneously with the addition of a single nonlytic reagent containing two peptide substrates. The live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant peptide substrate (GF-AFC Substrate). The substrate enters intact cells, where it is cleaved to generate a fluorescent signal proportional to the number of living cells. This live-cell protease activity marker becomes inactive upon loss of membrane integrity and leakage into the surrounding culture medium.

  A second, cell-impermeant, fluorogenic peptide substrate (bis-AAF-R110 Substrate) is used simultaneously to measure dead-cell protease activity that has been released from cells that have lost membrane integrity. This results in ratiometric, inversely correlated measures of cell viability and cytotoxicity. The ratio of viable cells to dead cells is independent of cell number and, therefore, can be used to normalize data. A second reagent containing luminogenic DEVD-peptide substrate for caspase-3/7 and Ultra-Glo™ Recombinant Thermostable Luciferase is added. Caspase-3/7 cleavage of the substrate releases luciferin, which is a substrate for luciferase and generates light. The light output, measured with a luminometer, correlates with caspase-3/7 activation as a key indicator of apoptosis.

  Cat.# G6320 contains sufficient reagents for 100 assays in a 96-well plate format or 400 assays in a 384-well format. Cat.# G6321 contains sufficient reagents for 500 assays in a 96-well plate format or 2,000 assays in a 384-well format.

• Protocols

  Complete Protocol

  

  ApoTox-Glo™ Triplex Assay Technical Manual

  PDF (630 KB)

  Quick Protocols

  

  ApoTox-Glo Triplex Assay Quick Protocol FB104

  PDF (110 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.

• Resources

  

  Articles

  • Add mix read assays for assessing cell health using an iPSC-derived cardiomyocyte cell model
  • Predictive Mechanism of Toxicity Using a Multiplexed Assay
  • Multiplexing Cell-Based Assays: Get More Biologically Relevant Data
  

  Citations

  • Cytotoxicity of propofol in human induced pluripotent stem cell-derived cardiomyocytes.
    --
    2018 J. Anesth.
  

  Software and Firmware

  • ApoToxGlo Triplex Assay Data Analysis Worksheet
  

  Other

  • Assays to Assess Cell Health Using an iPS Cardiomyocyte Cell Model Poster