Expression plasmid pCAD29 (pCAD29_hIRG1_4-461_pvp008, Addgene #124843)
E. coli BL21(DE3) CodonPlus-RIL competent cells
Kanamycin, 100 mg/ml in water
Chloramphenicol, 34 mg/ml in ethanol
SOC agar plates
Lysozyme
Biotin
Strep-Tactin XT superflow, 50% suspension (IBA GmbH)
YMC ECO15/200M0V or similar empty column
TEV protease
Syringe filter, 0.45 µm, Sartorius Minisart Plus
Vivaspin 20, MWCO 30,000 concentrator
High pressure homogeniser or sonifier. E.g. Avestin EmulsiFlex-C3 homogenizer or Bandelin Sonoplus 2000 Sonifier with a TT13Z probe
Äkta chromatography machine, 4 ml Strep-Tactin XT column, Superdex 200 26/60 column
SDS-PAGE equipmentAll buffers should be vacuum-filtered through a 0.45 µm filter.