TY  - JOUR
A1  - Brüchert, Stefan
A1  - Joest, Eike F.
A1  - Gatterdam, Karl
A1  - Tampé, Robert
T1  - Ultrafast in-gel detection by fluorescent super-chelator probes with HisQuick-PAGE
T2  - Communications biology
N2  - Polyacrylamide gel electrophoresis (PAGE) and immunoblotting (Western blotting) are the most common methods in life science. In conjunction with these methods, the polyhistidine-tag has proven to be a superb fusion tag for protein purification as well as specific protein detection by immunoblotting, which led to a vast amount of commercially available antibodies. Nevertheless, antibody batch-to-batch variations and nonspecific binding complicate the laborious procedure. The interaction principle applied for His-tagged protein purification by metal-affinity chromatography using N-nitrilotriacetic acid (NTA) was employed to develop small high-affinity lock-and-key molecules coupled to a fluorophore. These multivalent NTA probes allow specific detection of His-tagged proteins by fluorescence. Here, we report on HisQuick-PAGE as a fast and versatile immunoblot alternative, using such high-affinity fluorescent super-chelator probes. The procedure allows direct, fast, and ultra-sensitive in-gel detection and analysis of soluble proteins as well as intact membrane protein complexes and macromolecular ribonucleoprotein particles.
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/53157
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-531574
SN  - 2399-3642
VL  - 3
IS  - Article number: 138
PB  - Springer Nature
CY  - London
ER  -