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@ARTICLE{Kempe:826832,
      author       = {Kempe, Daryan and Cerminara, Michele and Poblete, Simón
                      and Schöne, Antonie and Gabba, Matteo and Fitter, Jörg},
      title        = {{S}ingle-{M}olecule {FRET} {M}easurements in
                      {A}dditive-{E}nriched {A}queous {S}olutions},
      journal      = {Analytical chemistry},
      volume       = {89},
      number       = {1},
      issn         = {1520-6882},
      address      = {Columbus, Ohio},
      publisher    = {American Chemical Society},
      reportid     = {FZJ-2017-01049},
      pages        = {694 - 702},
      year         = {2017},
      abstract     = {The addition of high amounts of chemical denaturants,
                      salts, viscosity enhancers or macro-molecular crowding
                      agents has an impact on the physical properties of buffer
                      solutions. Among others, the (microscopic) viscosity, the
                      refractive index, the dielectric constant, and the ionic
                      strength can be affected. Here, we systematically evaluate
                      the importance of solvent characteristics with respect to
                      single-molecule FRET (smFRET) data. First, we present a
                      confocal based method for the determination of fluorescence
                      quantum yields to facilitate a fast characterization of
                      smFRET-samples at sub-nM-concentrations. As a case study, we
                      analyze smFRET data of structurally rigid, double-stranded
                      DNA-oligonucleotides in aqueous buffer and in buffers with
                      specific amounts of glycerol, guanidine hydrochloride
                      (GdnHCl), and sodium chloride (NaCl) added. We show that the
                      calculation of interdye distances, without taking into
                      account solvent-induced spectral and photophysical changes
                      of the labels, leads to deviations of up to 4 Å from the
                      real interdye distances. Additionally, we demonstrate that
                      electrostatic dye–dye repulsions are negligible for the
                      interdye distance regime considered here (>50 Å). Finally,
                      we use our approach to validate the further compaction of
                      the already unfolded state of phosphoglycerate kinase (PGK)
                      with decreasing denaturant concentrations, a mechanism known
                      as coil–globule transition.},
      cin          = {ICS-5 / ICS-2},
      ddc          = {540},
      cid          = {I:(DE-Juel1)ICS-5-20110106 / I:(DE-Juel1)ICS-2-20110106},
      pnm          = {551 - Functional Macromolecules and Complexes (POF3-551)},
      pid          = {G:(DE-HGF)POF3-551},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000391346600055},
      doi          = {10.1021/acs.analchem.6b03147},
      url          = {https://juser.fz-juelich.de/record/826832},
}