Lewy body pathology in clinically unimpaired individuals is associated with worse smell and cognitive functions and predicted cognitive decline and progression to Parkinson’s disease or Lewy bodies dementia biofinder_study OskarHansson9
Purification of recombinant wild-type α-syn was performed as previously reportedBL21 bacteria from a glycerol stock were streaked on a selective plate containing kanamycin and incubated at 37 °C overnight. A single colony was selected and inoculated into 5 ml of Luria broth with kanamycin and allowed to grow for 4–5 h at 37 °C with continuous agitation at 250 rpm.
5 by the addition of 400–600 µl of NaOH 1 M . The protein extract was filtered through a 0.22 µm filter , loaded into a Ni–NTA column on an NGC chromatography system and washed with 20 mM Tris pH 7.5 at room temperature. The column was further washed with 50 mM imidazole in Tris 20 mM pH 7.5, generating a peak that was not collected. A linear gradient up to 500 mM imidazole in 20 mM Tris pH 7.5 was performed, and the peak collected between 30 and 75% of imidazole buffer .
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