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Renilla luciferase-labeled Annexin V: a new probe for detection of apoptotic cells.
- Source :
-
The Analyst [Analyst] 2012 Nov 07; Vol. 137 (21), pp. 5062-70. Date of Electronic Publication: 2012 Sep 24. - Publication Year :
- 2012
-
Abstract
- The Ca(2+)-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.
- Subjects :
- Antineoplastic Agents pharmacology
Humans
Jurkat Cells
Luciferases, Renilla genetics
Luciferases, Renilla isolation & purification
Molecular Probes genetics
Molecular Probes isolation & purification
Neutrophils cytology
Neutrophils drug effects
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins isolation & purification
Recombinant Fusion Proteins metabolism
Staining and Labeling
Annexin A5 metabolism
Apoptosis drug effects
Luciferases, Renilla metabolism
Molecular Probes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1364-5528
- Volume :
- 137
- Issue :
- 21
- Database :
- MEDLINE
- Journal :
- The Analyst
- Publication Type :
- Academic Journal
- Accession number :
- 23000925
- Full Text :
- https://doi.org/10.1039/c2an35741k