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. 2008 Dec 2;3(12):e3822. doi: 10.1371/journal.pone.0003822

Figure 3. Tandem Affinity Purification (TAP) of cytoplasmic dynein complexes using His-tag and SBP-tag.

Figure 3

mfGFP was fused to C-terminus of dynein intermediate chain (DIC). (A) Localization of DIC-mfGFP in living HeLa cells. Scale bar, 10 µm. (B) Isolation of cytoplasmic dynein complexes. Streptavidin (SA) column chromatography isolated DIC-mfGFP and dynein heavy chain (DHC) with some contaminating proteins in 50–75 kDa range (asterisk). Ni-NTA chromatography isolated the two proteins, but many contaminants were also seen. TAP strategy with Ni-NTA and SA (Ni-NTA + SA) isolated the two proteins with a significantly reduction of contaminants in 50–75 kDa range.