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Mitochondrial Isolation and Real-Time Monitoring of MOMP

  • Mark P. A. Luna-Vargas
  • Jarvier N. Mohammed
  • Jesse D. Gelles
  • Jerry Edward Chipuk
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1877)

Abstract

Isolated model systems have proven to be the standard in the apoptosis field to deconstruct MOMP into individual steps and to study the behavior of a subset of MOMP regulators. Here we describe the method to isolate, JC-1-label, and purify mouse liver mitochondria and subsequently describe how to utilize the JC-1-labeled mitochondria for real-time MOMP measurements.

Key words

Isolated mitochondria JC-1 labeling Real-time MOMP measurements BCL-2 family BAX 

Notes

Acknowledgments

We would like to thank everyone in the Chipuk Laboratory for their assistance and support. This work was supported by the following: NIH grants R01 CA157740 (J.E.C.) & R01 CA206005 (J.E.C.), the JJR Foundation, the William A. Spivak Fund, the Fridolin Charitable Trust, an American Cancer Society Research Scholar Award, a Leukemia & Lymphoma Society Career Development Award, and an Irma T. Hirschl/Monique Weill- Caulier Trust Research Award. This work was also supported in part by two research grants (5-FY11-74 and 1-FY13-416) from the March of Dimes Foundation, the Developmental Research Pilot Project Program within the Department of Oncological Sciences at the Icahn School of Medicine at Mount Sinai, and the Tisch Cancer Institute Cancer Center Support Grant (P30 CA196521).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Mark P. A. Luna-Vargas
    • 1
    • 2
  • Jarvier N. Mohammed
    • 1
    • 2
    • 3
  • Jesse D. Gelles
    • 1
    • 2
    • 3
  • Jerry Edward Chipuk
    • 1
    • 2
    • 3
    • 4
    • 5
  1. 1.Department of Oncological SciencesIcahn School of Medicine at Mount SinaiNew YorkUSA
  2. 2.The Tisch Cancer InstituteIcahn School of Medicine at Mount SinaiNew YorkUSA
  3. 3.The Graduate School of Biomedical SciencesIcahn School of Medicine at Mount SinaiNew YorkUSA
  4. 4.Department of DermatologyIcahn School of Medicine at Mount SinaiNew YorkUSA
  5. 5.The Diabetes, Obesity, and Metabolism InstituteIcahn School of Medicine at Mount SinaiNew YorkUSA

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