MitoSciences (MS1079)

Mitochondrial Viability Stain (ab129732)

Overview

  • Product nameMitochondrial Viability Stain
  • Detection methodFluorescent
  • Tests
    1 x 2000 test
  • Sample type
    Adherent cells, Suspension cells
  • Product overview

    ab129732 is a fluorometric/colorimetric assay that allows determination of the metabolic capacity of live cells in high throughput format. It uses an indicator dye to measure oxidation-reduction reactions which principally occur in the mitochondria of live cells. When reduced by metabolically active cells the non-fluorescent dark blue dye becomes fluorescent pink with absorbance at 570nm and red-fluorescent properties (560±10Ex/590Em) at neutral pH. The dye can be measured in fluorescence or absorbance mode. However fluorescence mode measurement offers greater assay linearity, reproducibility, robustness and sensitivity.

  • Tested applicationsFunctional Studiesmore details
  • PlatformReagents

Properties

  • Alternative names
    • Mitochondrial Viability Stain

Applications

Our Abpromise guarantee covers the use of ab129732 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent concentration.

Mitochondrial Viability Stain images

  • Effect of stain incubation time on assay reproducibility and background. Dark wall plates were seeded with a titration series of each cell line in a final volume of 100 µL of media. 2X stain (diluted in the cell line specific growth media) was added to each well at specified time points. XY graphs data is shown after media and background substraction. Bar graph shows mean background staining at different time points. 4 hour incubation of stain results in optimal reproducibility with minimal increase of background over the 2 hour incubation.
  • Mitochondrial viability stain in long term toxicity. HepG2 cells previously cultured in glucose or galactose substrate media were seeded at 10,000 cells per well and allowed to adhere overnight. Media was replaced for the specific culture media in the presence of a titration series of Rotenone (5 µM – 0.5 pM). Cells were treated for 72 hours prior to the addition of 2X stain. After 4 hours of incubation plates were analyzed by fluorescent readout.
  • Dynamic range and suitability of the mitochondrial viability assay for high throughput screening. Cells were seeded in a titration series with n=11 for each data point. Fluorometric readout gave on average a Z factor of 0.82 on Jurkat cells seeded from 1,500 – 1000,000 cells per well on a 96-well plate.
  • Dynamic range and suitability of the mitochondrial viability assay for high throughput screening. Cells were seeded in a titration series with n=11 for each data point. Colorimetric readout gave on average a Z factor of 0.72 on HepG2 cells seeded from 40,000 – 150,000 cells per well.
  • Jurkat cells were seeded at 25,000 cells per well in a 50 µL volume and immediately overlayed with a 2X concentration of Idarubicin for a final volume of 100 µL. Cells were incubated for 2 hours with Idarubicin prior to the addition of 2X stain diluted in RPMI media. After 4 hours of further incubation with stain, fluorescence was measured. IC50 for Idarubicin was found at 22 – 25 µM.
  • Jurkat cells were seeded at 25,000 cells per well in a 50 µL volume and immediately overlayed with a 2X concentration of Staurosporin for a final volume of 100 µL. Cells were incubated for 4 hours with Staurosporin prior to the addition of 2X stain diluted in RPMI media. After 4 hours of further incubation with stain, fluorescence was measured. IC50 for staurosporin was found at 500 nM.

Protocols

References for Mitochondrial Viability Stain (ab129732)

ab129732 has not yet been referenced specifically in any publications.

Product Wall

The colorimetric assay may be run on cell culture treated clear plastic plates.

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We only tested the cell lines shown in the graph, but the stain should work with any cell line (of any species).

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"