[ccp4bb] precipitates
Dear CCP4bbers, I am trying to crystallize a protein with a peptide attached to it via a 9aa linker. I am mostly getting precipitates in conditions carrying salts. I have tried optimization screens by varying the pH and salt concentrations, but in vain. Can anyone please suggest ways by which I could optimize the conditions further? thanks in advance, -- Amit Sharma
Re: [ccp4bb] precipitates
Dear Amit -- On 30 Sep 2008, at 14:16, amit sharma wrote: I am trying to crystallize a protein with a peptide attached to it via a 9aa linker. I am mostly getting precipitates in conditions carrying salts. I have tried optimization screens by varying the pH and salt concentrations, but in vain. Can anyone please suggest ways by which I could optimize the conditions further? Among all the possibilities, you can: - try to crystallize without salt ; - change the temperature ; - dilute your protein ; - add some isopropanol (or other) to make the solution "smoother" ; - change your linker size ; - ... HTH Kind regards. -- Leo -- Chavas Leonard, Ph.D. @ home Research Associate Marie Curie Actions Fellow Faculty of Life Sciences The University of Manchester The Michael Smith Building Oxford Road Manchester Lancashire M13 9PT Tel: +44(0)161-275-1586 e-mail: [EMAIL PROTECTED] http://personalpages.manchester.ac.uk/staff/leonard.chavas/
Re: [ccp4bb] precipitates
Dear All, Many thanks for the suggestions. Cheers, Amit 2008/10/1 Chavas Leo <[EMAIL PROTECTED]> > Dear Amit -- > On 30 Sep 2008, at 14:16, amit sharma wrote: > > I am trying to crystallize a protein with a peptide attached to it via a > 9aa linker. I am mostly getting precipitates in conditions carrying salts. I > have tried optimization screens by varying the pH and salt concentrations, > but in vain. Can anyone please suggest ways by which I could optimize the > conditions further? > > > Among all the possibilities, you can: > - try to crystallize without salt ; > - change the temperature ; > - dilute your protein ; > - add some isopropanol (or other) to make the solution "smoother" ; > - change your linker size ; > - ... > > HTH > Kind regards. > > -- Leo -- > > Chavas Leonard, Ph.D. @ home > Research Associate > Marie Curie Actions Fellow > > Faculty of Life Sciences > The University of Manchester > The Michael Smith Building > Oxford Road > Manchester Lancashire > M13 9PT > > Tel: +44(0)161-275-1586 > e-mail: [EMAIL PROTECTED] > http://personalpages.manchester.ac.uk/staff/leonard.chavas/ > > > -- Amit Sharma, Ph.D. Research Associate, Department of Biology, University of York, YO10 5DD UK
