[ccp4bb] How to get rid of Membrane formed on hanging droplets?

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: 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

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
---------------------
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 navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: MTZ format conversion
From: Kevin Cowtan cowtan {- at -} YSBL {- dot -} YORK {- dot -} AC {- dot -} UK
Date: 2007-03-05

Next message:
Subject: Re: protease cleavage sites
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2007-03-05