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.

Nazari M., Emamzadeh R., Hosseinkhani S., Cevenini L., Michelini E., Roda A. (2012). Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells. ANALYST, 137, 5062-5070 [10.1039/c2an35741k].

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

CEVENINI, LUCA;MICHELINI, ELISA;RODA, ALDO
2012

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.
2012
Nazari M., Emamzadeh R., Hosseinkhani S., Cevenini L., Michelini E., Roda A. (2012). Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells. ANALYST, 137, 5062-5070 [10.1039/c2an35741k].
Nazari M.; Emamzadeh R.; Hosseinkhani S.; Cevenini L.; Michelini E.; Roda A.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/130033
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