Mitochondrial Viability Stain (ab129732)

Overview

• Product nameMitochondrial Viability Stain
• 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 Studies more details

Properties

• Storage instructionsStore at +4°C. Please refer to protocols.

• Components	2000 tests
  20x Mitochondrial viability stain	1 x 24ml

• Research Areas
  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Cell Viability and Senescence Kits

  • Tags & Cell Markers
  • Subcellular Markers
  • Organelles
  • Mitochondria

Applications

Mitochondrial Viability Stain images

• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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.
• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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.
• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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.
• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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.
• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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.
• 

  Functional Studies - Mitochondrial Viability Stain (ab129732)
  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

Protocol Booklet