Hi, It's not very clear if your protein already crystallizes (maybe? otherwise why screen in a precise condition set such as yours?) and you're trying to get the ligand in...
The quasi-crystalline stuff in the wells looks to me like your compound is not very soluble in the condition(s) you work with and it is falling out of solution. It is a fairly common phenomenon in our (i.e. Pharma/Biotech) world where key research compounds often have aqueous solubility slightly worse than that of a Sony Walkman. You don't mention if your protein does the same when you set it up w/o compound so it could also be the protein of course. Note for the future - images are better as links, this way is kinder to people who read email on cellphones etc. Artem www.harkerbio.com "Crystallizing things, all kinds of things..." On Sat, Dec 17, 2016 at 3:01 AM, dhaval patel <pateldhaval...@gmail.com> wrote: > Dear CCP4 users, > > I am trying to co-crystal my protein of interest with a ligand and within > 2days of crystallization setup, I guess there is a formation of aggregrates > or something like that (see attachment picture). > > My experiment details are as follow: > Protein - 40kDa > LIgand- 1728 Da > Buffer condition - 25% PEG 1500 and its variation with no salts > Protein used for crystallization - 5mg/ml (130uM) in 20mM Tris pH 7.5 > Ligand used for crystallization - 250 uM, 500 uM and 1mM dissolved in > water. > Crystallization method - microbatch under oil > > I am observing the same kind of pattern in almost all wells in the plates. > > Any help is appreciated > > > > > -- > Dhaval Patel > PhD Student, > Bioinformatics & > Structural Biology > Indian Institute of > Advanced Research > +91-9925450504 <+91%2099254%2050504> > >
