It’s easy to do on a 0.2 µm spin filter, just pipet the crystals onto the filter with excess mother liquor, wash with several spins of mother-liquor, and elute with SDS loading buffer. Run all the washes and elution on a gel.
-- Kevin M. Jude, PhD Structural Biology Research Specialist, Garcia Lab Howard Hughes Medical Institute Stanford University School of Medicine Beckman B177, 279 Campus Drive, Stanford CA 94305 From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of MARCHOT Pascale <pascale.marc...@univ-amu.fr> Date: Wednesday, November 8, 2023 at 9:28 AM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: Re: [ccp4bb] What could these crystals be? ... and reveal by WB if the protein is tagged. The trickiest part of all this will be to rinse the "xtal(s)" as extensively as possible in a protein&DNA-free solution before dissolving it for SDS-PAGE/WB or MS or whichever method you select, in order to avoid contamination by the protein-DNA complex present in the drop solution. Paradoxically, the good news is that if the xtal collapses/melts easily during the rinsing steps, it will mean that it is protein. Best wishes - Pascale ________________________________ De : CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> de la part de Jurgen Bosch <jxb...@case.edu> Envoyé : mercredi 8 novembre 2023 18:08 À : CCP4BB@JISCMAIL.AC.UK Objet : Re: [ccp4bb] What could these crystals be? You could also run them on a gel if you don’t want to shot with canons on small birds. Jürgen On Nov 8, 2023, at 11:33 AM, M T <michel...@gmail.com> wrote: Dear Careina, If you have easy access to mass spectrometry, you can try to fish/rince your « pumpkin seeds » and send them to mass to try to identify what is inside to see if it needs optimization or not. Best. Le 8 nov. 2023 à 16:03, 000002531c126adf-dmarc-requ...@jiscmail.ac.uk a écrit : Hi all We have been trying with no success to crystalize a protein. Recently we got these strange shape "crystals". They are hard and flat but they do not diffract at all. Any ideas as to what could cause this? Careina ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 <IMG-20231108-WA0007.jpg> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
