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Re: [ccp4bb] Crystal Optimization |
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- Protein crystallographyMain steps:- Protein purification- Crystallisation Special:- Programs for crystallography- X-ray detectors Basic tutorials:- Chemistry- Protein - Peptide - Amino Acids Xtal community:- CCP4BB |
CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999Subject: Re: Crystal Optimization From: Muhammed bashir Khan muhammad {- dot -} bashir {- dot -} khan {- at -} UNIVIE {- dot -} AC {- dot -} AT Date: 2012-07-11 Dear All, Thanks alot for your valuable suggestion.I hope I will find out the solution now. As far as to giveup is out of question!!!! Thanks once agin Regards; Bashir On Wed, July 11, 2012 05:25, Tuhin Bhowmick wrote: > Dear Muhammad, > > I had a similar case, and the crystals could indeed be optimized. A few > things to check first, > > 1) How long does it take for the needles to appear? Sometimes, if the > protein is degraded/ cleaved over time, a small population (possibly a > fragment of the whole > protein) from the heterogeneous mix can give similar crystals. Like > Bryan had suggested, it is also very useful to check the mol wt. of the > crystallized species through > mass spect/ native page. But do make sure to give the crystals serial > washes, so the test accounts for crystallizexd species, not the ones from > surrounding condition. > > 2) If the crystals indeed contain the protein of interest, they can be > used > for various seeding methods. I've got results from both streak and micro > seeding. > > Best, > > Tuhin. > > Tuhin Bhowmick > > Department of Physics > > Indian Institute of Science > > Bangalore: 560012 > Email: tuhin.iisc@gmail.com > > On Wed, Jul 11, 2012 at 12:34 AM, Bernhard Rupp (Hofkristallrat a.D.) < > hofkristallamt@gmail.com> wrote: > >> > Always give up.**** >> >> ** ** >> >> ...definitely not the kind of guy I want to sit up front in an >> airliner…** >> ** >> >> ** ** >> >> Best regards, BR**** >> >> -----------------------------------------------------------------**** >> >> Bernhard Rupp, ATP-B737, CFII-MEI**** >> >> Vienna Air International**** >> >> Professional Aviation Services**** >> >> 001 (925) 209-7429**** >> >> +43 (676) 571-0536**** >> >> br@vienna-air.com**** >> >> br@ruppweb.org**** >> >> http://www.vienna-air.com/ **** >> >> -----------------------------------------------------------------**** >> >> It is not your aptitude but your attitude**** >> >> that determines your altitude. (or your crystals)**** >> >> -----------------------------------------------------------------**** >> >> ** ** >> >> ** ** >> >> ** ** >> >> From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of >> yybbll**** >> >> Sent: Tuesday, July 10, 2012 11:58 AM**** >> >> To: CCP4BB@JISCMAIL.AC.UK**** >> >> Subject: Re: [ccp4bb] Crystal Optimization**** >> >> ** ** >> >> Hi, **** >> >> **** >> >> In my experience, it is very very very difficult to optimize this needle >> like crystal. Always give up.**** >> >> **** >> >> Good luck!**** >> >> **** >> >> **** >> >> Dear All;**** >> >> Could somebody give a nice suggestion how the following type crystal >> could >> **** >> >> be optimized, I almost tried everything.**** >> >> Crystal Image is attached**** >> >> Crystal condition: 20% w/v PEG3350 and 200mM NaCl.**** >> >> Thanks in advance**** >> >> Bashir**** >> >> -- **** >> >> Muhammad Bashir Khan**** >> >> ****************************************************** >> >> Structural Genome Consortium (SGC). University of Toronto**** >> >> Toronto, Canada**** >> > -- Muhammad Bashir Khan ************************************************** Department for Structural and Computational Biology Max F. Perutz Laboratories University of Vienna Campus Vienna Biocenter 5 A-1030 Vienna Austria Austria Phone: +43(1)427752224 Fax: +43(1)42779522 CCP4bb navigationCCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999 |
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