Quantitation of apoptosis and necrosis by annexin V binding, propidium iodide uptake, and flow cytometry

Crowley, Lisa C., Marfell, Brooke J., Scott, Adrian P. and Waterhouse, Nigel J. (2016) Quantitation of apoptosis and necrosis by annexin V binding, propidium iodide uptake, and flow cytometry. Cold Spring Harbor Protocols, 2016 11: 953-957. doi:10.1101/pdb.prot087288


Author Crowley, Lisa C.
Marfell, Brooke J.
Scott, Adrian P.
Waterhouse, Nigel J.
Title Quantitation of apoptosis and necrosis by annexin V binding, propidium iodide uptake, and flow cytometry
Journal name Cold Spring Harbor Protocols   Check publisher's open access policy
ISSN 1559-6095
1940-3402
Publication date 2016-11-01
Sub-type Article (original research)
DOI 10.1101/pdb.prot087288
Open Access Status Not yet assessed
Volume 2016
Issue 11
Start page 953
End page 957
Total pages 5
Place of publication Cold Spring Harbor, NY, United States
Publisher Cold Spring Harbor Laboratory Press
Collection year 2017
Language eng
Abstract The surface of healthy cells is composed of lipids that are asymmetrically distributed on the inner and outer leaflet of the plasma membrane. One of these lipids, phosphatidylserine (PS), is normally restricted to the inner leaflet of the plasma membrane and is, therefore, only exposed to the cell cytoplasm. However, during apoptosis lipid asymmetry is lost and PS becomes exposed on the outer leaflet of the plasma membrane. Annexin V, a 36-kDa calcium-binding protein, binds to PS; therefore, fluorescently labeled Annexin V can be used to detect PS that is exposed on the outside of apoptotic cells. Annexin V can also stain necrotic cells because these cells have ruptured membranes that permit Annexin V to access the entire plasma membrane. However, apoptotic cells can be distinguished from necrotic cells by co-staining with propidium iodide (PI) because PI enters necrotic cells but is excluded from apoptotic cells. This protocol describes Annexin V binding and PI uptake followed by flow cytometry to detect and quantify apoptotic and necrotic cells.
Q-Index Code CX
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Mater Research Institute-UQ (MRI-UQ)
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Created: Tue, 15 Nov 2016, 00:51:22 EST by Julia McCabe on behalf of Learning and Research Services (UQ Library)