| X-RAY DIFFRACTION |
8.5 |
293.15 |
Samples were centrifuged at 13,000 x g for 3 min to remove macro-aggregates and dust, and crystallised inside an anaerobic chamber (N2/H2 (97:3%) atmosphere, 20 degrees Celsius). Crystallisation was done by the sitting drop method in 96-Well MRC 2-Drop polystyrene Crystallisation Plates (SWISSCI) plate containing 90 uL of crystallisation solution in the reservoir in all cases. Crystals were obtained by mixing 0.7 uL of crystallisation solution with 0.7 uL of protein sample at a concentration of 2.95 mg/mL. The crystallisation solution contained the following: 30 % v/v 2-methyl-2,4-pentanediol, 100 mM Tris pH 8.5, 500 mM Sodium chloride, and 8 % w/v Polyethylene glycol 8,000 (Crystallisation solution of the JBScreen Wizard form Jena Bioscience, Germany). |
