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Flow Cytometry-Based Detection and Analysis of BCL-2 Family Proteins and Mitochondrial Outer Membrane Permeabilization (MOMP)

  • Lindsey M. Ludwig
  • Katrina L. Maxcy
  • James L. LaBelle
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1877)

Abstract

The BCL-2 family of proteins orchestrates a complex signaling network that governs the balance between cellular survival and death. A comprehensive understanding of the mechanistic interactions between these proteins continues to evolve in normal and malignant cells. The functional variation by individual BCL-2 proteins in different cell types has driven clinical therapeutic development in targeting individual BCL-2 members with the goal of fine-tuning cell death in diseased cells. Given the importance of understanding and validating the effect of activating or inhibiting BCL-2 protein interactions in individual cells, the methods used to measure apoptotic cell death have undergone increased scrutiny. Here, we describe two in vitro flow cytometry-based methods that are useful in measuring BCL-2 proteins and mitochondrial-based cell death in complex cell populations.

Key words

BCL-2 proteins Intracellular staining Flow cytometry Mitochondria outer membrane permeabilization (MOMP) Apoptosis 

Notes

Acknowledgment

We would like to thank Eric E. Smith for graphics in Fig. 2 and 5.

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Copyright information

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

Authors and Affiliations

  • Lindsey M. Ludwig
    • 1
  • Katrina L. Maxcy
    • 1
  • James L. LaBelle
    • 1
  1. 1.Section of Hematology/Oncology/Stem Cell Transplantation and Committee on Cancer Biology, Department of PediatricsUniversity of ChicagoChicagoUSA

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