Hi Dilip- Try using your microcrystals as seeds and try the matrix microseeding method:
Acta Crystallogr D Biol Crystallogr.<javascript:AL_get(this,%20'jour',%20'Acta%20Crystallogr%20D%20Biol%20Crystallogr.');>2007 Apr;63(Pt 4):550-4. Epub 2007 Mar 16. An automated microseed matrix-screening method for protein crystallization. D'Arcy A<http://www.ncbi.nlm.nih.gov/pubmed?term=%22D%27Arcy%20A%22%5BAuthor%5D>, Villard F<http://www.ncbi.nlm.nih.gov/pubmed?term=%22Villard%20F%22%5BAuthor%5D>, Marsh M<http://www.ncbi.nlm.nih.gov/pubmed?term=%22Marsh%20M%22%5BAuthor%5D> . Or, consider the alternative explanation: *Acta Cryst.* (2008). D*64*, 1222-1227 [ doi:10.1107/S0907444908031302<http://dx.doi.org/10.1107/S0907444908031302>] The role of bias in crystallization conditions in automated microseedingF. J. St John<http://scripts.iucr.org/cgi-bin/citedin?search_on=name&author_name=St%20John,%20F.J.>, B. Feng<http://scripts.iucr.org/cgi-bin/citedin?search_on=name&author_name=Feng,%20B.>and E. Pozharski<http://scripts.iucr.org/cgi-bin/citedin?search_on=name&author_name=Pozharski,%20E.> For some reason, it sometimes works. Laurie On Sun, Oct 10, 2010 at 1:15 AM, Dilip Badgujar <[email protected]>wrote: > hi guys > > I am trying to crystallize protein complexed with peptide(14mer). > In initial screening i got microcrystals in following condition > - 0.2M MgCL2 > 0.1M Tris pH 8.5 > 28 % PEG 4000 > Ptotein concentration - 12.5mg/ml and peptide conc.-5mg/ml > Complex ratio- (Protein-Peptide)(1:1.5) > Incubation temp.-22 degree > Method - Sitting drop vapor diffusion method > > within four hours of setting trials I can see too many microcrystals. > Though i tried reduction of protein and precipitant conc. as well as using > mineral oil to decrease evaporation rate but still it is not working. > > I am waiting for your valuable suggestions. > > <http://sigads.rediff.com/RealMedia/ads/click_nx.ads/www.rediffmail.com/signatureline....@middle?>
