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Detection of MLKL Oligomerization During Programmed Necrosis

  • Zhenyu Cai
  • Zheng-Gang Liu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1857)

Abstract

Programmed necrosis, also known as necroptosis, is a form of regulated necrotic cell death that is mediated by receptor-interacting protein kinases RIP1 (or RIPK1), RIP3 (or RIPK3), and the mixed lineage kinase domain-like protein, MLKL. Following the induction of programmed necrosis, MLKL is phosphorylated by RIP3 and oligomerizes and then the protein translocates to cell plasma membrane in order to execute programmed necrosis. Here, we describe a detailed protocol to detect MLKL oligomerization in necroptotic cells by Western blotting analysis under nonreducing condition. Therefore, we established the method to detect the activation of programmed necrotic pathway.

Key words

MLKL Necroptosis Nonreducing SDS-PAGE Immunoblots 

Notes

Acknowledgments

This research was supported by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Center for Cancer Research, National Cancer InstituteNational Institutes of HealthBethesdaUSA
  2. 2.National Center for Liver CancerShanghaiChina

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