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Promega Corporation

Mitochondrial Toxicity Assay

The Mitochondrial ToxGlo™ Assay is a cell-based assay method that employs a sequential addition, multiplexed assay chemistry for predicting potential mitochondrial dysfunction as a result of xenobiotic exposure. The assay is based on the differential measurement of biomarkers associated with changes in cell membrane integrity and cellular ATP levels relative to vehicle-treated control cells during short exposure periods. Cell membrane integrity is first assessed by measuring the presence or abscence of a distinct protease activity associated with necrosis using a fluorogenic peptide substrate (bis-AAF-R110) to mea...

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Mitochondrial ToxGlo™ Assay

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Mitochondrial ToxGlo™ Assay

 Components

  • bis-AAF-R110 Substrate

    G609A1 x 10μl
  • Assay Buffer

    G610A1 x 10ml
  • ATP Detection Buffer

    G806A1 x 10ml
  • ATP Detection Substrate

    V363A1 x 1 vial
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  • Helix Freezer

This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.


Click here to learn more about the Helix Collection »


Already a Helix Customer?

Request to add this product to your Helix »


10ml
- G8000 $ 199.00 Add to cart

Mitochondrial ToxGlo™ Assay

Close Window

Mitochondrial ToxGlo™ Assay

 Components

  • bis-AAF-R110 Substrate

    G609B2 x 50μl
  • Assay Buffer

    G610B1 x 50ml
  • ATP Detection Buffer

    G806B1 x 100ml
  • ATP Detection Substrate

    V363B1 x 1 vial
Close Window
  • This product can be added to a

  • Helix Freezer

This product is available through the Promega Helix onsite stocking program in a –20°C Helix Freezer. The program offers numerous convenient solutions to meet your lab's needs. Helix Freezers are available in two sizes: 5.7 cubic ft. or 9.7 cubic ft.


Click here to learn more about the Helix Collection »


Already a Helix Customer?

Request to add this product to your Helix »


100ml
- G8001 $ 935.00 Add to cart


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Storage Conditions

Store the Mitochondrial ToxGlo™ Assay components at –20°C.

For product intended use please see Patents & Disclaimers tab.

Schematic of the Mitochondrial ToxGlo™ Assay.Figure 1. Schematic of the Mitochondrial ToxGlo™ Assay.
Mitochondrial responsiveness to a model mitochondrial toxin in the presence of galactose or glucose.Figure 2. Mitochondrial responsiveness to a model mitochondrial toxin in the presence of galactose or glucose.

Some cells treated in the presence of glucose may preferentially rely on glycolysis to meet bioenergetic needs and are therefore relatively unresponsive to mitochondrial toxins (Glucose ATP). Cells treated in the presence of galactose must use oxidative phosphorylation to generate ATP and are therefore more responsive to mitochondrial perturbation (Galactose ATP). Oligomycin treatment did not cause changes in membrane integrity in either formulation of medium (Galactose Cytotoxicity and Glucose Cytotoxicity). Shown are data from K562 cells at 10,000 cells/well in a white Costar® 96-well plate. The cells were exposed to oligomycin for 2 hours.

Representative profiles of mitochondrial toxicity with the Mitochondrial ToxGlo™ Assay.Figure 3. Representative profiles of mitochondrial toxicity with the Mitochondrial ToxGlo™ Assay.

K562 cells were plated at 10,000 cells/well in white 96-well plates (Costar®) and treated with serial dilutions of compounds resuspended in glucose-free (galactose-supplemented) RPMI 1640 media for
2 hours. Panel A shows no changes in ATP or membrane integrity (MI), which indicates that the compound is not a mitochondrial toxin. Panel B. The reduction in ATP with commensurate MI changes indicate that the compound is not a mitochondrial toxin; instead primary necrosis is taking place. Panel C. The reduction in ATP with no changes in MI indicates that the compound is a mitochondrial toxin. Panel D. The reduction in ATP with discordant changes in MI indicate that the compound is a mitochondrial toxin. Note: If a decrease in fluorescence, or both fluorescence and luminescence, are observed it is typically due to color quenching interferences adversely affecting assay measures. If the cells are dosed in glucose-containing medium, compounds producing ATP-depletion effects should be counter-screened in galactose-containing medium to rule out inhibition of glycolysis.

K562 cells were plated at 10,000 cells/well in white 96-well plates (Costar®) and treated with serial dilutions of compounds resuspended in glucose-free (galactose-supplemented) RPMI 1640 media for
2 hours. Panel A shows no changes in ATP or membrane integrity (MI), which indicates that the compound is not a mitochondrial toxin. Panel B. The reduction in ATP with commensurate MI changes indicate that the compound is not a mitochondrial toxin; instead primary necrosis is taking place. Panel C. The reduction in ATP with no changes in MI indicates that the compound is a mitochondrial toxin. Panel D. The reduction in ATP with discordant changes in MI indicate that the compound is a mitochondrial toxin. Note: If a decrease in fluorescence, or both fluorescence and luminescence, are observed it is typically due to color quenching interferences adversely affecting assay measures. If the cells are dosed in glucose-containing medium, compounds producing ATP-depletion effects should be counter-screened in galactose-containing medium to rule out inhibition of glycolysis.

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Use Restrictions

G8000, G8001 For Research Use Only. Not for Use in Diagnostic Procedures.

Patents - Disclaimers

G8000, G8001 U.S. Pat. Nos. 7,083,911, 7,452,663 and 7,732,128 and other patents.

G8000, G8001 U.S. Pat. Nos. 6,602,677, 7,241,584 and 8,030,017 and other patents and patents pending.

G8000, G8001 The method of recombinant expression of Coleoptera luciferase is covered by U.S. Pat. Nos. 5,583,024, 5,674,713 and 5,700,673.

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