hey! I would like some more information... which buffers have you used and
at what pH?

Em qui., 9 de jan. de 2025, 22:01, Marco Bravo <
0000d0eb7bee83ae-dmarc-requ...@jiscmail.ac.uk> escreveu:

> 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
>
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