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ApoTox-Glo™ Triplex Assay

Assess Viability, Cytotoxicity and Apoptosis in One Assay

  • Easy to implement with an "add-mix-measure" protocol
  • Normalize data with a built-in control
  • Flexible and easily automated

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Catalog number selected: G6320

$ 597.00
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ApoTox-Glo™ Triplex Assay
10ml
$ 597.00
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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.

    Measure Viability, Cytotoxicity, and Apoptosis in the Same Sample Well

    Necrosis Results

    Necrosis Results
    Necrosis Results

    Ionomycin treatment of Jurkat cells for 6 hours results in dose-dependent decrease in viability and increase in cytotoxicity with no caspase-3/7 activation, consistent with primary necrosis.

    Apoptosis Results

    Apoptosis Results
    Apoptosis Results

    Staurosporine treatment of Jurkat cells for 6 hours results in a dose-dependent decrease in viability and increase in cytotoxicity with an increase in caspase-3/7 activity consistent with apoptosis.

    Cytostasis Results

    Cytostasis Results
    Cytostasis Results

    Camptothecin treatment of Jurkat cells for 48 hours results in dose-dependent decrease in viability, no cytotoxicity and increase in caspase-3/7 activity, consistent with cell-cycle arrest and early-phase apoptosis.

    Specifications

    You are viewing: G6320 Change Configuration

    What's in the box?

    Item Part # Size

    GF-AFC Substrate

    G608A 1 × 10μl

    bis-AAF-R110 Substrate

    G609A 1 × 10μl

    Assay Buffer

    G610A 1 × 10ml

    Caspase-Glo® 3/7 Buffer

    G810A 1 × 10ml

    Caspase-Glo® 3/7 Substrate

    G811A 1 × 1 bottle

    SDS

    Choose language:

    Download SDSPDF (216 KB) – English (United States)

    Certificate of Analysis

    See all certificates for G6320

    Use Restrictions

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

    Storage Conditions

    BB

    Patents and Disclaimers

    European Pat. No. 1131441 and Japanese Pat. No. 4520084.

    U.S. Pat. Nos. 7,416,854, 7,553,632 and other patents pending.

    Specifications

    You are viewing: G6321 Change Configuration

    What's in the box?

    Item Part # Size

    GF-AFC Substrate

    G608B 1 × 50μl

    bis-AAF-R110 Substrate

    G609B 1 × 50μl

    Assay Buffer

    G610A 2 × 10ml

    Caspase-Glo® 3/7 Buffer

    G810A 5 × 10ml

    Caspase-Glo® 3/7 Substrate

    G811A 5 × 1 bottle

    SDS

    Choose language:

    Download SDSPDF (220 KB) – English (United States)

    Certificate of Analysis

    See all certificates for G6321

    Use Restrictions

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

    Storage Conditions

    BB

    Patents and Disclaimers

    European Pat. No. 1131441 and Japanese Pat. No. 4520084.

    U.S. Pat. Nos. 7,416,854, 7,553,632 and other patents pending.

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