Hi Marco, I wonder what is the highest concentration of the protein you've tried. Once I worked with a protein which would crystallize only if it was concentrated into a jell... Good luck! Nukri
On Thu, Jan 9, 2025 at 7:01 PM Marco Bravo < 0000d0eb7bee83ae-dmarc-requ...@jiscmail.ac.uk> wrote: > Hello all, > I am seeking some advice or ideas to crystalize a nuclease protein. It > exists within a heterodimer, homodimer, and maybe monomer as well depending > on the species from which the protein is in. The protein has been shown to > have nuclease activity on its own depending on the species. I am screening > the protein from three different species and have setup so many crystal > high-throughput crystal screens with and without DNA. I never get protein > crystals for these proteins, I have done thermal shift assays to identify a > buffer that stabilizes the protein better than the storage buffer and found > some conditions that increased the melting temp by 4-5 degrees. I screened > the protein again in these new stabilizing buffers but still have not > gotten any hits. There are protein structures for the protein in the pdb > with its partner protein in the heterodimer formation. I have experience > screening other proteins and getting several hits per kit so I know what to > expect for a successful screen. Any advice would be helpful, I want to > retain the full-length protein for structural studies. I would appreciate > any ideas or advice for moving forward and trying to obtain crystals. > Thanks! > > Marco > > ######################################################################## > > 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/ > ######################################################################## 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/
