Relatives discovered: Membraneproteins of cyanobacteria and higher organisms are structurally highly similar uni_mainz_eng
2 cells were used to inoculate 1 L LB medium , and cells were grown at 37 °C until an ODDLP was induced via the addition of IPTG . Cells were grown at 20 °C overnight, harvested by centrifugation the following day, and the resulting pellet was snap frozen in liquid nitrogen and stored at −20 °C until used.
After incubation at 4 °C for 2 h, the matrix was washed six times with the same buffer supplemented with either 20, 40, or 50 mM imidazole, respectively.DLP was incubated with 20 mM DTT for half an hour at 4 °C and the protein was further purified via size exclusion chromatography using a self-packed Sephacryl S-400 HR column equilibrated with 20 mM HEPES pH 7.4, 0.2 mM DTT on an ÄKTA purifier 10 system . Main peak fractions were collected and concentrated.
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