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Mitochondrial Isolation and Real-Time Monitoring of MOMP

Mark P A Luna-Vargas1,2, Jarvier N Mohammed1,2,3, Jesse D Gelles1,2,3

  • 1Department of Oncological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 12, 2018
PubMed

Insights

This study details a method for isolating and labeling mouse liver mitochondria using JC-1 dye. This technique enables real-time measurement of mitochondrial outer membrane permeabilization (MOMP) in apoptosis research.

Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Isolated model systems are standard for studying apoptosis.
  • These systems deconstruct mitochondrial outer membrane permeabilization (MOMP) into steps.
  • Studying MOMP regulators requires robust experimental models.

Purpose of the Study:

  • To describe a method for isolating mouse liver mitochondria.
  • To detail the process of labeling these mitochondria with JC-1.
  • To enable real-time measurements of MOMP using prepared mitochondria.

Main Methods:

  • Isolation of mitochondria from mouse liver.
  • Labeling of isolated mitochondria with the JC-1 dye.
  • Purification of JC-1-labeled mitochondria.
  • Utilizing labeled mitochondria for real-time MOMP assays.

Main Results:

  • Successful isolation and purification of mouse liver mitochondria.
  • Effective labeling of mitochondria with JC-1.
  • Demonstration of JC-1-labeled mitochondria for real-time MOMP monitoring.

Conclusions:

  • The described method provides a reliable approach for studying MOMP.
  • This technique facilitates real-time analysis of apoptosis progression.
  • The protocol is valuable for investigating MOMP regulators in isolated mitochondria.
Keywords:
BAXBCL-2 familyIsolated mitochondriaJC-1 labelingReal-time MOMP measurements

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