Inducing apoptosis using chemical treatment and acidic pH, and detecting it using the Annexin V flow cytometric assay

PLoS One. 2022 Jun 29;17(6):e0270599. doi: 10.1371/journal.pone.0270599. eCollection 2022.

Abstract

Cell death is important in physiology, and can happen as a result of structural damage, or as a sequence of programmed cellular processes known as apoptosis. Pathogenic alterations in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune diseases, immunodeficiencies, and degenerative conditions. Developing accurate and reproducible laboratory methods for inducing and detecting apoptosis is vital for research into these conditions. A number of methods are employed to detect cell death, including DNA fragmentation, the TUNEL assay, and electron microscopy although each has its limitations. Flow cytometry allows for the distinction between live, early apoptotic, late apoptotic and necrotic cells. In this protocol we successfully induce apoptosis using chemical treatment and treatment with low pH in solid tumour cell lines, and have optimized detection using the Annexin V/PI apoptosis assay.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Annexin A5 / metabolism
  • Apoptosis*
  • Flow Cytometry / methods
  • Humans
  • Hydrogen-Ion Concentration
  • Necrosis

Substances

  • Annexin A5

Grants and funding

This research was funded by The National Research Foundation (NRF) Thuthuka program (www.nrf.ac.za), grant number TTK20110727000022309. This was awarded to CM Worsley. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.