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Issue 21, 2012
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Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells

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Abstract

The Ca2+-dependent binding of Annexin V to phosphatidylserine on cell surfaces is a reliable marker for apoptosis that is widely used in flow cytometry based apoptosis assays. In this paper, we report a new class of Annexin V-based probes for apoptosis. Luciferase from Renilla reniformis (RLuc) was linked to Annexin V and expressed successfully in a soluble form in Escherichia coli BL21 (DE3). The new probe, Rluc/Annexin V, was purified and functionally assayed for detection of apoptosis in actinomycin D-induced apoptotic Jurkat cells. Moreover, the spontaneous apoptosis in neutrophils was shown using the new probe. The results indicate that Rluc/Annexin V can bind to the apoptotic cells, and the signal of Renilla luciferase can be detected by luminometric measurements. The availability of Rluc/Annexin V may be of potential commercial interest for improving current apoptosis assays.

Graphical abstract: Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells

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Supplementary files

Article information


Submitted
04 Jun 2012
Accepted
31 Aug 2012
First published
24 Sep 2012

Analyst, 2012,137, 5062-5070
Article type
Paper

Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells

M. Nazari, R. Emamzadeh, S. Hosseinkhani, L. Cevenini, E. Michelini and A. Roda, Analyst, 2012, 137, 5062
DOI: 10.1039/C2AN35741K

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