PS I would also try substituting imidazole for HEPES and zinc sulphate for zinc chloride and sodium sulphate.
Mark J van Raaij Laboratorio M-4 Dpto de Estructura de Macromoleculas Centro Nacional de Biotecnologia - CSIC c/Darwin 3 E-28049 Madrid, Spain tel. (+34) 91 585 4616 http://www.cnb.csic.es/content/research/macromolecular/mvraaij On 27 Sep 2011, at 18:02, Mark J van Raaij wrote: > Hi Christopher, > you'll have to try to find out how they mix the components at MD and if they > pH the solution afterwards or before (or not at all...) and to which pH. You > can ask them; or try different mixing/pHing protocols on a small scale and > see if one works in keeping all in solution. > Mark > > Mark J van Raaij > Laboratorio M-4 > Dpto de Estructura de Macromoleculas > Centro Nacional de Biotecnologia - CSIC > c/Darwin 3 > E-28049 Madrid, Spain > tel. (+34) 91 585 4616 > http://www.cnb.csic.es/content/research/macromolecular/mvraaij > > > > > > On 27 Sep 2011, at 17:55, Browning Christopher wrote: > >> Dear All, >> >> My question might be a bit out of place, but perhaps someone can help. I've >> screened my protein in the Nuc-Pro screen from Molecular Dimensions and >> found some crystals in condition B10. They seem to be protein crystals as >> they are not highly optically active and look different to salt crystals. So >> condition B10 consist of 10% PEG 4K, 50mM Imidazole pH 7.2, 20mM Zinc >> sulfate. In the screen, this condition is perfectly clear, but when I try >> and make my own screen, the whole solution turns white. Apparently, Zinc >> sulfate/Imidazole can be used for staining SDS-gels, but that does not >> really help my a lot. Does anybody have an idea how I might be able to keep >> everything in solution, seeing that the MD guys managed somehow...... I'm a >> bit desperate as I don't have many hits for this protein. >> >> Cheers, >> >> Chris B >> >> -- >> Dr. Christopher Browning >> Post-Doctor to Prof. Petr Leiman >> EPFL >> BSP-416 >> 1015 Lausanne >> Tel: 0041 (0) 02 16 93 04 40
