Mitochondrial Isolation and Real-Time Monitoring of MOMP
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 wordsIsolated mitochondria JC-1 labeling Real-time MOMP measurements BCL-2 family BAX
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).