Re: [ccp4bb] Protein aggregation and crystallization

2011-08-28 Thread Allan Pang
Hi Anita, Won't harm if you put it on crystallization tray. You never know what these proteins might do. Allan Quoting Zheng Zhou : Hi, Anita If you could find a way to test the elute's activity/binding to its' substrat/cofactor, then you will learn much more about your target. If the fun

[ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread Allan Pang
Hi there everyone, What does it mean when you have proteins eluting in almost the whole column volume of S200? I ran a gel with fractions from 8ml to 20ml and saw band for my protein all throughout. Judging peaks on chromatogram is not useful as it doesn't have any aromatic rings. Che

[ccp4bb] Hiro Tsuruta

2011-08-28 Thread Tsutomu Matsui
It is with great sadness that we announce that Dr. Hirotsugu (Hiro) Tsuruta, a senior scientist and biophysicist in the Structural Molecular Biology program at Stanford Synchrotron Radiation Lightsource (SSRL) at SLAC, passed away on August 25, 2011, in Japan, after a courageous fight with canc

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread Roger Rowlett
It possibly means: 1. Your sample application volume was too large 2. Total protein quantity was too large 3. Fraction size was too large 4. Sample was too crude (GEC is a finishing step, not a starting step for protein purification. 5. Sample was too concentrated/viscous 6. Flow rate was too fast

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread David Briggs
Following on from Roger's fine suggestions: 8. Your column is knackered. Can you see fine lines or cracks in the column? Good packing is v.important for SEC columns. HTH, Dave David C. Briggs PhD Father, Structural Biologist and Sceptic

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread Nian Huang
Hi, David, What is the common cause of knackered SEC column? Will equilibrizing a buffer containing 150 mM NaCl directly into a 20% EtOH or vise versa cause the problem. There was no problem just after packing the column. Nian On Sun, Aug 28, 2011 at 9:24 AM, David Briggs wrote: > Following on f

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread David Briggs
Hi Nian, It can be a number of things - but typically, air getting into the column, or a leaky seal somewhere letting it dry out. Switching from 150mM NaCl to 20% EtOH directly shouldn't cause a problem. The reason I suggest it is that I have had the same situation as described in the original p

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread Filip Van Petegem
worst-case scenario for crystallization purposes: 9. Your protein runs as a mixture of monomers, dimers, trimers, whatever-mers... Filip On Sun, Aug 28, 2011 at 7:24 AM, David Briggs wrote: > Following on from Roger's fine suggestions: > > 8. Your column is knackered. Can you see fine lines or

[ccp4bb] phaser problem

2011-08-28 Thread Amir Khan
Hi Has anyone a solution to the following problem with Phaser (version 2.1.4)? UNHANDLED ERROR: basic_string::_S_create It seems to be data and model independent – there was an old thread from earlier this year, but I cannot find a follow-up. Thanks! Amir

Re: [ccp4bb] Protein elution in Size Exclusion

2011-08-28 Thread Mario Sanches
Testing for cracks in the column is quite easy. Take whichever standard you happen to have (lisozyme, GST, etc) and do a run. If it comes out in a single peak the problem is with your prep. If it comes out throughout the run the problem is with your column. Mario Sanches On Sun, Aug 28, 2011 at 5

[ccp4bb] Windows 7 and Xtal Software

2011-08-28 Thread Jacob Keller
Dear Crystallographers, are there any additional problems or known issues running ccp4 or other xtal software on windows 7 (beyond those of Vista, etc.?) Your input would be really appreciate before I sink my own personal $$$ into a new laptop Jacob Keller -- *

Re: [ccp4bb] Methods for dehydrating crystals

2011-08-28 Thread Matthew BOWLER
Dear Andrea, as others have suggested there are many ways to dehydrate crystals by increasing the concentration of salt/precipitant or adding glycerol/EG/PEG 200-400 to the solutions surrounding your crystals. I have always found controlled dehydration using a specific device to be much

[ccp4bb] Self-rotation Patterson maps

2011-08-28 Thread john peter
Hello, I wish to learn about self-rotation Patterson maps such as how to interpret a map and find out non-crystallographic symmetry. Could you suggest some good papers (for dummies), web-sites etc. Thanks a lot for your help. John

Re: [ccp4bb] HTP ligand screening

2011-08-28 Thread Alexander . Schiffer
Dear Jacob, Globalphasing has a new programm called Pipedream that should do just what you want. I am not sure if it is available to non consortium members. Best regards, Alexander -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Jacob Keller Sen

Re: [ccp4bb] Windows 7 and Xtal Software

2011-08-28 Thread Nat Echols
On Sun, Aug 28, 2011 at 7:23 PM, Jacob Keller < j-kell...@fsm.northwestern.edu> wrote: > are there any additional problems or known issues running ccp4 or > other xtal software on windows 7 (beyond those of Vista, etc.?) Phenix, ARP/wARP, and HKL2000 do not run on Windows. I'm pretty sure none