Fig. 5: Selective protein detection in SDS-PAGE and native PAGE by fluorescent chelator probes.
From: Ultrafast in-gel detection by fluorescent super-chelator probes with HisQuick-PAGE
a, b Detection of His12-tagged reference proteins (300 ng, 7 pmol) in cell lysates of bacterial and various eukaryotic origins (2.5 µg total protein) by hexaNTAAlexa647 (a) or immunoblotting using two different primary antibodies (b). *, nonspecific band. c Detection of a purified His10-tagged membrane protein complex (TmrAB, 1.3 µg, 500 nM) in two different commonly used membrane mimetics (DDM and MSP1D1 nanodiscs) were tested on a blue native PAGE by tris- and hexaNTAAlexa647 (450 nM). d Visualization of protein complexes containing a His6-tagged component on native PAGE. Non-tagged 30S ribosomal subunit (8 pmol) with fourfold molar equivalence of ABCE1-His6 (32 pmol, 2.3 µg) was used to verify the formation of the 30S·ABCE1-His6 post-splitting complex. Due to the low amount of ABCE1-His6, the His-tagged recycling factor was not visualized by Coomassie staining15. e Direct contrast of denaturating HisQuick-PAGE using hexaNTAAlexa647 (450 nM) and 3xFLAG immunoblot detection using various amounts of purified 3xFLAG- and His10-tagged reference protein (nanobody C-terminally fused to 3xFLAG- and His10-tag).
