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Figure 4
Setting up an in meso crystallization trial manually involves (a) placing membrane-protein solution and lipid into separate gas-tight micro-syringes (typically 50 or 100 µl) connected by a narrow-bore coupler, (b) passing the protein solution and lipid from one syringe to the other via the coupler to effect mixing, homogenization and spontaneous self-assembly of the cubic phase into the bilayer of which the protein has become reconstituted, (c) transferring the optically clear mesophase into one of the syringes, (d) replacing the empty syringe with a dispensing micro-syringe (typically 10 µl) mounted in a repeat dispenser and transferring protein-laden mesophase from the large to the small syringe by way of the coupler, (e) dispensing mesophase followed by precipitant solution into the wells of a glass sandwich crystallization plate and (f) sealing the wells with a glass cover slide. The remaining wells on the plate are filled and sealed and the plate is then incubated at the desired temperature to allow crystallization to occur. An open-access online video of the entire procedure is available (Caffrey & Porter, 2010BB24).

Journal logoSTRUCTURAL BIOLOGY
COMMUNICATIONS
ISSN: 2053-230X
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