Dear All,
I wonder if someone could give an advice on LCP crystallisation? In particular- we have a problem with mixing the protein with oil in gas-tight
coupled syringes. We used monoolein (Sigma), and tried to act according to the protocol in (Aherne et al, Appl Crystallogr. 2012). We warmed up both monoolein and the protein, had no problem loading the melted oil in the syringe, both components looked nice and transparent in separate syringes, but after we joined them and start mixing, the mixture soon became white as chalk, and even if mixed during 20 munutes, remained opaque. We used 3:2 oil to protein ratio, tried several proteins, with and without detergents, several mixing times, and it still is white.
We still set up a couple of trays anyway (on Mosquito robot, this part worked fine) - but what should have been the mesophase looked wrong.
Would greatly appreciate an advice on proper mixing! Or on good oils, or any other tips.
Many thanks,
Olga
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