Issue 21, 2012
From the journal:

Analyst

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

Mahboobeh Nazari,a   Rahman Emamzadeh,*b   Saman Hosseinkhani,c   Luca Cevenini,d   Elisa Michelinid  and  Aldo Rodad  

* Corresponding authors

a Department of Recombinant Technology Research, Nanobiotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

b Department of Biology, Faculty of Science, University of Isfahan, Isfahan, Iran
E-mail: Sci_rahman@yahoo.com, r.emamzadeh@sci.ui.ac.ir
Fax: +98 311 6682910
Tel: + 98 311 7932174

c Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

d Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy

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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Article information

DOI
https://doi.org/10.1039/C2AN35741K
Article type
Paper
Submitted
04 Jun 2012
Accepted
31 Aug 2012
First published
24 Sep 2012

Analyst, 2012,137, 5062-5070

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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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