Dear Yahui, I also suggest In-situ dehydration combined with in-situ diffraction . See paper below "Using high-throughput in situ plate screening to evaluate the effect of dehydration on protein crystals", Alice Douangamath et al, Acta Cryst. (2013). D69, 920–923
------------------------------------------------------------ Dr Isabel De Moraes, MRSC Membrane Protein Laboratory Diamond Light Source ------------------------------------------------------------ On 22 Apr 2014, at 13:44, Campeotto, Ivan wrote: Dear Yahui, Your problem is quite common and very frustrating, I know. 1) Did you try to find new crystallisation conditions by re-screening using micro-seeds? 2) Did you try Additive Screens? 3) In-situ diffraction may also help you to have a feeling about the diffraction power of your crystals in a sealed enviroment at RT. This strategy depends on how many crystals you have and how many you are willing to sacrifice. 4) One of the last resorts would be to go back to cloning and either generate new protein constructs of the same proteins or move towards protein homologues of one of both proteins. This strategy is of course very time consuming but it may pay you back in the long distance. Also a nice paper, which I would recommend is: "A review of techniques for maximizing diffraction from a protein crystal in stilla". Janet Newman, Acta Cryst. (2006). D62, 27–31 Good luck! Dr Ivan Campeotto Imperial College London On 22 Apr 2014, at 12:24, Yahui Liu wrote: Dear all, Right now, we are try to crystallise an protein complex. By all kinds of efforts, the resolution of the crystal was about 8 angstrom. So we would like to try dehydration. The precipitant that our crystal grew was ethanol. Since the evaporation of the ethanol, our crystal is unstable at the room temperature or 4 degree when it was exposed to the air directly. Do any people have some suggestions about the dehydration way of our crystal? Best Yahui -- This e-mail and any attachments may contain confidential, copyright and or privileged material, and are for the use of the intended addressee only. If you are not the intended addressee or an authorised recipient of the addressee please notify us of receipt by returning the e-mail and do not use, copy, retain, distribute or disclose the information in or attached to the e-mail. Any opinions expressed within this e-mail are those of the individual and not necessarily of Diamond Light Source Ltd. Diamond Light Source Ltd. cannot guarantee that this e-mail or any attachments are free from viruses and we cannot accept liability for any damage which you may sustain as a result of software viruses which may be transmitted in or with the message. Diamond Light Source Limited (company no. 4375679). Registered in England and Wales with its registered office at Diamond House, Harwell Science and Innovation Campus, Didcot, Oxfordshire, OX11 0DE, United Kingdom
