(1) PlasmidConstruction and Protein Expression
Creative Biostructure is specialized in plasmid construction of full-length, site-mutated, truncated and tagged proteins. The constructed gene is cloned into a suitable vector and expressed in an appropriate cell line.
(2) Purification of Crystallization-grade Proteins
Proteins are purified using Akta (GE Healthcare) with multiple chromatographic methods including ion exchange, affinity, size exclusion, etc. Resins with specific affinity to the tags are used in the case of tagged proteins. Protein purity is checked via SDS-PAGE and western blots.
(3) Initial Crystallization Screening
Hundreds of non-redundant crystallization conditions will be screened by taking advantage of the high-throughput crystallization facility alongside multiple crystallization approaches, such as vapor diffusion crystallization, seeding and co-crystallization.