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TU Berlin

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Publikationen 2005 - 2009

Gold Fluorescent Annexin A5 as a Novel Apoptosis Detection Tool
Zitatschlüssel 061.2009.kurschus
Autor Kurschus, F. and Pal, P. P. and Bäumler, P. and Jenne, D. E. and Wiltschi, B. and Budisa, N.
Seiten 626-633
Jahr 2009
DOI 10.1002/cyto.a.20737
Journal Cytometry Part A
Jahrgang 75
Nummer 7
Zusammenfassung 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.
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