Annexin V-Affinity assay: A review on an apoptosis detection system based on phosphatidylserine exposure

Abstract

Apoptosis is a programmed, physiological mode of cell death that plays an important role in tissue homeostasis. Understanding of the basic mechanisms that underlie apoptosis will point to potentially new targets of therapeutic treatment of diseases that show an imbalance between cell proliferation and cell loss. In order to conduct such research, techniques and tools to reliably identify and enumerate death by apoptosis are essential. This review focuses on a novel technique to detect apoptosis by targeting for the loss of phospholipid asymmetry of the plasma membrane. It was recently shown that loss of plasma membrane asymmetry is an early event in apoptosis, independent of the cell type, resulting in the exposure of phosphatidylserine (PS) residues at the outer plasma membrane leaflet. Annexin V was shown to interact strongly and specifically with PS and can be used to detect apoptosis by targeting for the loss of plasma membrane asymmetry. Labeled annexin V can be applied both in flow cytometry and in light microscopy in both vital and fixed material by using appropriate protocols. The annexin V method is an extension to the current available methods. This review describes the basic mechanisms underlying the loss of membrane asymmetry during apoptosis and discusses the novel annexin V‐binding assay. Cytometry 31:1–9, 1998. © 1998 Wiley‐Liss, Inc.

Early loss of mammalian target of rapamycin complex 1 (mTORC1) signalling and reduction in cell size during dominant-negative suppression of hepatic nuclear factor 1-α (HNF1A) function in INS-1 insulinoma cells

“…5). Annexin V detects the exposure of phosphatidylserine on the outer plasma membrane, a hallmark of apoptosis [28]. In agreement with our previous data [12,13], we detected elevated levels of Annexin V-positive apoptotic cells from 36 h onwards (Fig.…”

In vitro measurement of cell death with the annexin A5 affinity assay

“…Green fluorescent annexin A5 is the most widely used target-specific probe for measurement of cell death. The annexin A5 affinity assay discriminates among living cells, cells in the early phase of cell death and (secondary) necrotic cells that have a compromised cell membrane 3,4 .…”

“…PS expressed at the cell surface can be measured by flow cytometry and by confocal scanning-laser microscopy (CSLM) using the PS-binding protein annexin A5 conjugated to fluorochromes. In conjunction with the permeability probe propidium iodide, a distinction can be made between dying cells with intact plasma-membrane integrity and (secondary) necrotic cells 3,4 .…”

Caspase-3 triggers early synaptic dysfunction in a mouse model of Alzheimer's disease

“…The exposure of phosphatidylserine (PS) residue on the plasma membrane is usually considered as a downstream event of caspase-3 activation and a hallmark of apoptosis 26 . In order to examine apoptosis in hippocampal synapses, we analyzed the exposure of PS in the synaptosomal fraction where active caspase-3 accumulates.…”