To identify non-specific biotinylation events, HEK293 cells transfected with a plasmid carrying only the biotin ligase moiety were used as negative controls to BioID experiments. Cells were lysed in the presence of detergents. After purification of biotinylated proteins on streptavidin-conjugated beads, the eluates were precipitated with TCA. After washing with acetone, the protein mixtures were digested with endoproteinase Lys-C and trypsin (Promega) and analyzed by MudPIT.
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: exogenous biotin ligase ; HEK293 cells ; BioID ; negative control
Principal Investigators: (in alphabetical order) |
Laurence Florens, The Stowers Institute for Medical Research, USA |
Submitting User: | laflorens |
Redwine WB, DeSantis ME, Hollyer I, Htet ZM, Tran PT, Swanson SK, Florens L, Washburn MP, Reck-Peterson SL.
The human cytoplasmic dynein interactome reveals novel activators of motility.
Elife. Epub 2017 Jul 18.
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Experimental Design | ||
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