Journal: Turkish Journal of Biology
Article Title: A tetracycline-inducible Split TurboID system for specific biotinylation and identification of nuclear proteins from HEK293T cells
doi: 10.55730/1300-0152.2734
Figure Lengend Snippet: (A) Western blot analysis to assess biotinylation activity of Split TurboID in tetracycline-induced and rapamycin-added cultures. Cell-free extracts were prepared from HEK-293 TetR+ cells to serve as the negative control. (B) Western blot analysis to demonstrate the efficiency of protein enrichment using streptavidin conjugated magnetic beads. (C) Western blot and SDS-PAGE analyses of the enrichment process. The membrane was probed with HRP-conjugated streptavidin to visualize biotinylated proteins. A replica SDS-PAGE gel was run and stained with silver to visualize proteins. Lane numbers: 1, Wash fraction 1; 2, Wash fraction 2; 3, Elution fraction 1; 4, magnetic beads used in the enrichment process. Nonbiotinylated samples that were also subjected to the enrichment process were from a whole cell-lysate that were prepared from nonbiotinylated cells.
Article Snippet: The cells were then incubated with Texas Red conjugated Streptavidin- (Thermo Fisher Sci, USA) for 1 h at room temperature.
Techniques: Western Blot, Activity Assay, Negative Control, Protein Enrichment, Magnetic Beads, SDS Page, Membrane, Staining