Dear everyone,
I am working on crystallization of a protein/RNA complex recently. The crystals were initially grown from BICINE(9.0) 0.1M, 1,4-Dioxane 2%(v/v) , PEG 20,000 10%(w/v), at 10mg/ml. I noticed that there was a membrane formed on the surface of the hanging droplets. This membrane seems very sticky. Consequently, almost all of the crystals stick to this membrane and can't be seperated for data collection. Sitting drops were also tried but crystals stick to the bottom of the sitting well. Different buffer(Tris, CHES), different PEG(PEG 8000, PEG3350, from 10%-1%) and different protein concentration (10-3mg/ml) were also tried, but the sticky membrane was still there. Does anyone have some experience solving this problem? Any suggestions would be highly appreciated!
Yeming
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Yeming Wang, Ph.D.
Laboratory of Structural Biology: Macromolecular Structure Group
National Institute of Environmental Health Sciences
National Institute of Health
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