He hanging-drop vapor diffusion method at 293 K, using a Mosquito nanodrop
169.00 Rs.
Published date: 2024-11-14
- Location: 16225, Haryana, India
He hanging-drop vapor diffusion method at 293 K, using a Mosquito nanodrop dispenser (TTP Labtech, Cambridge, MA, USA), followed by manual optimization. In the initial screening, 200 nl aliquots of the protein solution were mixed with an equal volume of reservoir solutions from commercially available screening kits, JCSG Core 1 to 4 (Qiagen, Chuo, Tokyo, Japan). TkoGINS crystallized under nine conditions out of the 384 in the semi-automatic setup. An optimization procedure yielded the best diffraction quality crystals, which were obtained by mixing 1 l of the protein solution with an equal volume of a reservoir, containing 0.1 M 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (pH 7.5), 0.2 M magnesium chloride and 24 to approximately 26 (v/v) PEG400. The crystals belonged to the space group P4 1 2 1 2, with unit cell dimensions a = b = 74.3 ? c = 181.3 ? and contained one Gins51 subunit and one Gins23 subunit per asymmetric unit.
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