Journal
/
Biology
/
Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells
JoVE Journal
Biology
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biology
Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

Measuring Mitochondrial Substrate Flux in Recombinant Perfringolysin O-Permeabilized Cells

DOI:
10.3791/62902-v

•

06:17 min

•

August 13, 2021

•

, , , , ,
1Department of Physiology, Faculty of Medicine in Hradec Králové,Charles University, 2Department of Pathophysiology, Third Faculty of Medicine,Charles University, 3Department of Internal Medicine,University Hospital Kralovske Vinohrady

Chapters

  • 00:04Introduction
  • 01:03Day 1
  • 03:00Day 2
  • 04:57Results: Mitochondrial Respiration Measurement
  • 05:33Conclusion

Summary

Automatic Translation

Automatic Translation

In this work, we describe a modified protocol to test mitochondrial respiratory substrate flux using recombinant perfringolysin O in combination with microplate-based respirometry. With this protocol, we show how metformin affects mitochondrial respiration of two different tumor cell lines.

Tags

Keywords: Mitochondrial Substrate FluxRecombinant Perfringolysin O-permeabilized CellsCancer Cell ResponseMetformin TreatmentMitochondrial MetabolismSeahorse XF 96 Cell Culture MicroplateOligomycinFCCPRotenone Antimycin A

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/62902?article_type=v

A Method For Production of Recombinant mCD1d Protein in Insect Cells.
check_url/62902?article_type=v

Probing for Mitochondrial Complex Activity in Human Embryonic Stem Cells
check_url/62902?article_type=v

Measuring the Osmotic Water Permeability Coefficient (Pf) of Spherical Cells: Isolated Plant Protoplasts as an Example
check_url/62902?article_type=v

Measuring Glutathione-induced Feeding Response in Hydra
check_url/62902?article_type=v

Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
check_url/62902?article_type=v

Assessing Autophagic Flux by Measuring LC3, p62, and LAMP1 Co-localization Using Multispectral Imaging Flow Cytometry
check_url/62902?article_type=v

A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions
check_url/62902?article_type=v

Transient Expression and Cellular Localization of Recombinant Proteins in Cultured Insect Cells
check_url/62902?article_type=v

Measurement of Energy Metabolism in Explanted Retinal Tissue Using Extracellular Flux Analysis
check_url/62902?article_type=v

Measuring Diurnal Rhythms in Autophagic and Proteasomal Flux

Read Article