Hi all- I interpreted the original question, wrongly so, as an inquiry into using cryoEM for 3D *crystallography, *which is very new, and not general 3D structure determination, which of course has been utilized for the last few decades. My apologies! Yes, there are plenty of success stories out there for using cryoEM to solve 3D structures, especially recent efforts with single particle analysis and the use of direct detectors.
Indeed, in my experience, you only need a few uL of a modest concentration to perform cryoEM experiments; however, size, shape, symmetry, and dispersity of the sample as well as buffer composition are major variables in whether you have success or not. Good luck! Brad On Fri, Aug 29, 2014 at 11:06 AM, Andy Sen <[email protected]> wrote: > Dear Rana, > > Brad's post compelled me to reply to you. The Cryo EM field is well into > its pro stage. > > If you can be specific about the MolWt, size and if it makes any complex, > then much precise and helpful answers to your question can be provided. > > Best > > Anindito Sen Ph.D > JeoL > > > On 29/08/2014, at 11:31 pm, Marcus Fislage <[email protected]> wrote: > > > > Hi Rana, > > > > in contrast to Brads answer, and in case I understand you well, Cryo EM > is not in ts infancy stage (as long as you do not talk about Cryo EM on > crystals. > > For cryo EM you need only several ul of a nM solution. > > I can always suggest you the 3dem bb. There are of course still > limitataions on the size of protein target and resolution, but those are > slowly falling. > > > > Some reading material > > http://www.ncbi.nlm.nih.gov/pubmed/23181775 > > http://www.ncbi.nlm.nih.gov/pubmed/25071206 > > > > > > Cheers > > Marcus > > > > Quoting Brad Bennett <[email protected]>: > > > >> Yes, but the technique is still in its infancy stage. > >> > >> DOI: 10.1016/j.sbi.2014.03.004 > >> > >> Cheers- > >> Brad > >> > >> > >> On Fri, Aug 29, 2014 at 6:34 AM, rana ibd < > >> [email protected]> wrote: > >> > >>> Dear CCP4 > >>> I wanted to ask has anyone tested 3D protein structure by cryo-electron > >>> microscopy? what is the suitable concentration required for this > procedure > >>> Best Regards > >>> Rana > > > > > > > > -- > > Marcus Fislage, PhD > > > > Howard Hughes Medical Institute (HHMI) > > Columbia University > > Department of Biochemistry and Biophysics > > Lab of Joachim Frank > > New York, NY > > > > Phone: 212.305.9524 > > Fax: 212.305.9500 >
