Open AccessOne-step Split GFP Staining for Sensitive Protein Detection and Localization in Mammalian Cells
Author(s) -
Lara Kaddoum,
Eddy Magdeleine,
Geoffrey S. Waldo,
Etienne Joly,
Stéphanie Cabantous
Publication year - 2010
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000113512
Subject(s) - green fluorescent protein , flow cytometry , microbiology and biotechnology , staining , protein subcellular localization prediction , epitope , recombinant dna , subcellular localization , biology , protein fragment complementation assay , fluorescence microscope , transfection , fluorescence , antibody , chemistry , complementation , biochemistry , cytoplasm , phenotype , gene , genetics , physics , quantum mechanics
Although epitope tags are useful to detect intracellular proteins and follow their localization with antibodies, background and nonspecific staining often remain problematic. We describe a simple assay based on the split GFP complementation system. Proteins tagged with the 15-amino acid GFP 11 fragment are detected with a solution of the recombinant nonfluorescent complementary GFP 1-10 fragment to reconstitute a fluorescent GFP. In contrast to antibody-based staining methods, this one-step assay presents high specificity and very low background of fluorescence, thus conferring higher signal-to-noise ratios. We demonstrate that this new application of the split GFP tagging system facilitates detection of proteins displaying various subcellular localizations using flow cytometry and microscopy analysis.
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