Annexin V-FITC Apoptosis Detection Kit validated via the IncuCyte S3 and OperettaCLS automated imaging platforms

INTRODUCTION

Annexins are a family of calcium-dependent phospholipid-binding proteins. They are abundant in eukaryotic organisms and belong to a family of ubiquitous cytoplasmic proteins involved in signal transduction. As a member of the annexin family, Annexin V stands out for its unique ability to bind selectively to phosphatidylserine (PS), a phospholipid typically found in the inner leaflet of the cell membrane. During apoptosis, specific cellular pathways are activated that expose PS to the cell surface, which has been shown to modulate immune responses by inhibiting procoagulant and proinflammatory activities in these dying cells. This binding to PS at the cell membrane has made Annexin V an invaluable tool in biomedical research, particularly in the fields of cell biology, immunology, and oncology. Annexin V interaction with PS has been exploited for imaging techniques surrounding apoptosis detection.
Enzo’s Annexin V-FITC Apoptosis Detection Kit (ALX-850-020) enables the classification and quantification of apoptotic cells by flow cytometry, fluorescence microscopy, as well as other means of fluorescent detection. FITC labeling allows for simple direct detection, and additional counterstaining by propidium iodide allows for the discrimination of apoptotic and necrotic cells with compromised cell membranes. Standard flow cytometry is a commonly used detection method. Still, it does not allow for extended analysis of cells over time or the production of composite images of the cells of interest.
Here, Enzo’s Annexin V-FITC Apoptosis Detection Kit is used to analyze carcinoma, glioblastoma, and fibroblast cell lines over time through fluorescent imaging analysis on the IncuCyte® S3 and Operetta CLS™ automated platforms from Sartorius and Perkin Elmer, respectively. Ultimately, this demonstrates the versatility of applying Enzo’s range of live cell dyes for quantifying apoptosis in various cultured cells on innovative imaging platforms.

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