- Having trouble crystallizing your protein/nucleic acid/complex?
- Getting crystals, but they don’t diffract well? Or do you require a different morphology or space group?
YES & YES?
Then take part in the Activity A1: Bring your sample at the HTCC6, organized by Croatian Association of Crystallographers and Douglas Instruments! If you have a moment, complete the online form before you come, or just bring your sample to the workshop.
As for the sample, we’ll need roughly 10 – 60 µL of solution, usually at around 10 mg/mL (although the best concentration depends on the solubility of the sample). If you happen to already have those poor crystals, and/or crystals of a structurally related macromolecule, just bring them along, that’d be awesome. If not, bring the solution, we’ll manage.
Students will be taught how to make seed-stocks and given guidance for setting up crystallization experiments. They will also be shown how to set up microbatch-under-oil experiments, which are easier to scale up to make samples for serial data-collection, neutron diffraction, cryoEM etc.
Important: We won’t be able to fit in samples from all delegates so that the number will be limited to approximately 10 samples. To guarantee your place, please complete and submit the online sample questionnaire.
SESSIONS:
- 07/04/2024, 21:00: Students who have brought samples should approach Douglas Instruments at the A1 desk to discuss their projects.
- 08/04/2024, 21:00: Short theoretical introduction on the sample preparation for advanced methods of structural determination, followed by practical demonstration of how to make seed stock and set up microseeding experiments.
- 10/04/2024, 14:00: Some of the new crystals obtained will be shown.
- 11/04/2024, 18:30: Wrap-up (a short look back at the activity within the closing ceremony)
The practical sessions will be coordinated by Douglas Instruments who are kindly making equipment available for the workshop.
REFERENCES:
[1] D’Arcy, Allan, Frederic Villard, and May Marsh. “An automated microseed matrix-screening method for protein crystallization.” Acta Crystallographica Section D: Biological Crystallography 63.4 (2007): 550-554.
[2] Shaw Stewart, Patrick D., et al. “Random microseeding: a theoretical and practical exploration of seed stability and seeding techniques for successful protein crystallization.” Crystal Growth & Design 11.8 (2011): 3432-3441.
[3] Abuhammad, Areej, et al. “Structure of arylamine N-acetyltransferase from Mycobacterium tuberculosis determined by cross-seeding with the homologous protein from M. marinum: triumph over adversity.” Acta Crystallographica Section D: Biological Crystallography 69.8 (2013): 1433-1446.
