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Detection of Necroptosis by Phospho-RIPK3 Immunohistochemical Labeling

Joshua D Webster1, Margaret Solon1, Susan Haller1

  • 1Department of Pathology, Genentech, South San Francisco, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 24, 2018
PubMed

Insights

This study introduces a new method using antibodies to detect autophosphorylated receptor interacting protein kinase 3 (RIPK3) in mouse tissues. This technique reliably identifies cells undergoing necroptosis, a form of programmed cell death.

Area of Science:

  • Cell Biology
  • Immunology
  • Pathology

Background:

  • Necroptosis is a regulated form of necrosis crucial in immunity and disease.
  • Receptor interacting protein kinase 3 (RIPK3) activation is a key event initiating necroptosis.
  • RIPK3 phosphorylates mixed lineage kinase-like (MLKL), leading to cell membrane damage and lysis.

Purpose of the Study:

  • To develop and validate an immunohistochemical method for detecting necroptosis in mouse tissues.
  • To utilize antibodies targeting RIPK3 autophosphorylation as a specific marker for necroptotic cells.

Main Methods:

  • Immunohistochemistry was employed to label autophosphorylated mouse RIPK3.
  • The method was applied to detect necroptotic cells within mouse tissue samples.

Main Results:

  • Successful development of an immunohistochemical assay for detecting RIPK3 autophosphorylation.
  • Demonstrated the utility of this assay in identifying necroptotic cells in mouse tissues.

Conclusions:

  • Immunohistochemical labeling of autophosphorylated RIPK3 provides a reliable method for monitoring necroptosis induction in vivo.
  • This technique offers a valuable tool for studying the role of necroptosis in various biological contexts and diseases.
Keywords:
AutophosphorylationImmunohistochemistryMouseNecroptosisRIPK3

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