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A bright and photostable fluorescent dye with a disulfide (S−S) linker and maleimide group (Rho594‐S2‐mal), as cleavable and reactive sites, was synthesized and conjugated with anti‐GFP nanobodies (NB). The binding… Click to show full abstract
A bright and photostable fluorescent dye with a disulfide (S−S) linker and maleimide group (Rho594‐S2‐mal), as cleavable and reactive sites, was synthesized and conjugated with anti‐GFP nanobodies (NB). The binding of EGFP (FRET donor) with anti‐GFP NB labeled with one or two Rho594‐S2‐mal residues was studied in vitro and in cellulo. The linker was cleaved with dithiothreitol recovering the donor (FP) signal. The bioconjugates (FP‐NB‐dye) were applied in FRET‐FLIM assays, confocal imaging, and superresolution STED microscopy.
Journal Title: Chembiochem
Year Published: 2022
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