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[ccp4bb] How to get rid of Membrane formed on hanging droplets? |
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CCP4bb navigationCCP4bb <-- 2007 <-- March 2007 <-- 05 March 2007Subject: How to get rid of Membrane formed on hanging droplets? From: "Wang, Yeming (NIH/NIEHS) [F]" WangY2 {- at -} NIEHS {- dot -} NIH {- dot -} GOV Date: 2007-03-05 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 --------------------- Yeming Wang, Ph.D. Laboratory of Structural Biology: Macromolecular Structure Group National Institute of Environmental Health Sciences National Institute of Health Mailing Address: Street Address: NIEHS, MD F3-05 NIEHS, Building 101, Room F363 P.O. BOX 12233 111 T.W. Alexander Drive RTP, NC 27709 RTP, NC 27709 Tel (o): 919-316-4634 E-mail: wangy2@niehs.nih.gov CCP4bb navigationCCP4bb <-- 2007 <-- March 2007 <-- 05 March 2007 |
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