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Gold fluorescent annexin A5 as a novel apoptosis detection tool
Author(s) -
Kurschus Florian C.,
Pal Prajna Paramita,
Bäumler Petra,
Jenne Dieter E.,
Wiltschi Birgit,
Budisa Nediljko
Publication year - 2009
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.20737
Subject(s) - annexin a5 , fluorescence , annexin , fluorescein isothiocyanate , flow cytometry , phosphatidylserine , biophysics , chemistry , fluorescein , förster resonance energy transfer , apoptosis , microbiology and biotechnology , biology , membrane , biochemistry , optics , phospholipid , physics
We describe a golden fluorescent apoptosis detection tool, which we generated by a fusion of golden fluorescent protein (GdFP) with human annexin A5 (anxA5). GdFP was obtained by replacement of tryptophan at position 66 with 4‐aminotryptophan in the chromophore of enhanced cyan fluorescent protein. The GdFP‐anxA5 construct combines highly desirable features originating from both fusion partners. These include (i) strong binding to membrane phosphatidylserine patches of apoptotic cells in the presence of Ca 2+ which is brought about by anxA5, (ii) the stable and homogeneous monomeric state, (iii) as well as the red‐shifted fluorescence maximum at 574 nm originating from GdFP. We found that GdFP‐anxA5 is equally well applicable for apoptosis studies as a routinely used fluorescein 5′‐isothiocyanate‐annexin A5 conjugate. Golden fluorescent annexin A5 represents a new, stable, and homogeneous red‐shifted optical probe for the efficient detection of apoptosis by fluorescence microscopy or by flow cytometry. © 2009 International Society for Advancement of Cytometry