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Colorful protein-based fluorescent probes for collagen imaging

PLoS ONE, 2014, Vol.9(12), pp.e114983-1/21 [Peer Reviewed Journal]

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  • Title:
    Colorful protein-based fluorescent probes for collagen imaging
  • Author: Aper, S.J.A. ; Spreeuwel, van, A.C.C. ; Turnhout, van, M.C. ; van der Linden, A.J. ; Pieters, P.A. ; Zon, van der, N.L.L. ; Rambelje, de la, S.L. ; Bouten, C.V.C. ; Merkx, M.
  • Contributor: Protein Engineering ; Soft Tissue Engineering and Mechanobiology ; Department of Biomedical Engineering
  • Found In: PLoS ONE, 2014, Vol.9(12), pp.e114983-1/21 [Peer Reviewed Journal]
  • Subjects: Journal Article ; Research Support, Non-U.S. Gov'T
  • Language: English
  • Description: Real-time visualization of collagen is important in studies on tissue formation and remodeling in the research fields of developmental biology and tissue engineering. Our group has previously reported on a fluorescent probe for the specific imaging of collagen in live tissue in situ, consisting of the native collagen binding protein CNA35 labeled with fluorescent dye Oregon Green 488 (CNA35-OG488). The CNA35-OG488 probe has become widely used for collagen imaging. To allow for the use of CNA35-based probes in a broader range of applications, we here present a toolbox of six genetically-encoded collagen probes which are fusions of CNA35 to fluorescent proteins that span the visible spectrum: mTurquoise2, EGFP, mAmetrine, LSSmOrange, tdTomato and mCherry. While CNA35-OG488 requires a chemical conjugation step for labeling with the fluorescent dye, these protein-based probes can be easily produced in high yields by expression in E. coli and purified in one step using Ni2+-affinity chromatography....
  • Identifier: ISSN: ; ISSN: 1932-6203

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