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@ARTICLE{Tashev:292129,
      author       = {S. A. Tashev and J. Euchner and K. Yserentant and S.
                      Hänselmann and F. Hild and W. Chmielewicz and J. Hummert
                      and F. Schwörer$^*$ and N. Tsopoulidis and S. Germer and Z.
                      Saßmannshausen and O. T. Fackler and U. Klingmüller$^*$
                      and D.-P. Herten},
      title        = {{P}ro{DOL}: a general method to determine the degree of
                      labeling for staining optimization and molecular counting.},
      journal      = {Nature methods},
      volume       = {21},
      number       = {9},
      issn         = {1548-7091},
      address      = {London [u.a.]},
      publisher    = {Nature Publishing Group},
      reportid     = {DKFZ-2024-01624},
      pages        = {1708-1715},
      year         = {2024},
      note         = {#LA:B200# / 2024 Sep;21(9):1708-1715},
      abstract     = {Determining the label to target ratio, also known as the
                      degree of labeling (DOL), is crucial for quantitative
                      fluorescence microscopy and a high DOL with minimal
                      unspecific labeling is beneficial for fluorescence
                      microscopy in general. Yet robust, versatile and easy-to-use
                      tools for measuring cell-specific labeling efficiencies are
                      not available. Here we present a DOL determination technique
                      named protein-tag DOL (ProDOL), which enables fast
                      quantification and optimization of protein-tag labeling.
                      With ProDOL various factors affecting labeling efficiency,
                      including substrate type, incubation time and concentration,
                      as well as sample fixation and cell type can be easily
                      assessed. We applied ProDOL to investigate how human
                      immunodeficiency virus-1 pathogenesis factor Nef modulates
                      CD4 T cell activation measuring total and activated copy
                      numbers of the adapter protein SLP-76 in signaling
                      microclusters. ProDOL proved to be a versatile and robust
                      tool for labeling calibration, enabling determination of
                      labeling efficiencies, optimization of strategies and
                      quantification of protein stoichiometry.},
      cin          = {B200},
      ddc          = {610},
      cid          = {I:(DE-He78)B200-20160331},
      pnm          = {312 - Funktionelle und strukturelle Genomforschung
                      (POF4-312)},
      pid          = {G:(DE-HGF)POF4-312},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:39117875},
      doi          = {10.1038/s41592-024-02376-6},
      url          = {https://inrepo02.dkfz.de/record/292129},
}