| X-RAY DIFFRACTION |
7.5 |
293.15 |
The crystal was obtained from an initial screening using the sitting drop method performed on a 96-well MRC 2-well polystyrene crystallisation plate (SWISSCI, Switzerland) at 20 degree Celsius under a pure N2 atmosphere. The plate was filled with 90 uL crystallisation solution in the large reservoir and the purified protein solution concentrated to 4.4 mg/mL was spotted in the small reservoirs by mixing 0.5 uL purified protein with 0.5 uL crystallisation solution using an OryxNano robot (Douglas Instruments Ltd, UK). The crystallisation solution was composed of: 20 % v/v Jeffamine M-600 pH 7.0, and 100 mM HEPES pH 7.5.
Once prepared, the plate was transferred to an anaerobic chamber containing an N2/H2 (97/3 percent) atmosphere for long-term storage.
Prior liquid nitrogen freezing, the crystal was soaked in the crystallization solution supplemented with 25 % v/v glycerol for a few seconds. |
