The NKI Protein Facility is a protein production, crystallization and biophysical characterization facility at the Division of Biochemistry at The Netherlands Cancer Institute (NKI) in Amsterdam.
The Protein Facility of the Netherlands Cancer Institute (NKI) in Amsterdam has a strong record in structural biology using macromolecular X-ray crystallography, Electron Microscopy and sample preparation, and is also extremely well-equipped for studying molecular biophysics.
With respect to Molecular Biophysics we offer to our users fluorescence-based fast kinetics and high-throughput experiments, ITC, MST, MALS, SPR and Thermal Shift Assays.
The Protein Facility of the Netherlands Cancer Institute (NKI) in Amsterdam has a strong record in structural biology using macromolecular X-ray crystallography, Electron Microscopy and sample preparation, and is also extremely well-equipped for studying molecular biophysics.
We are very experienced in macromolecular crystallization and offer to our users:
- Robotics for preparation of nanoliter crystallization drops in 96-well format.
- Storage of crystallization plates at 4°C or 20°C.
- Screening and ranking of crystallization conditions in 96-well format (TOPAZ, AutoInspex station).
Solving crystal structures is beyond the scope of the facility!
The Protein Facility of the Netherlands Cancer Institute (NKI) in Amsterdam has a strong record in structural biology using macromolecular X-ray crystallography, Electron Microscopy and sample preparation, and is also extremely well-equipped for studying molecular biophysics.
We are very experienced in protein production using insect and mammalian cells, and assist users with the design of their expression construct in terms of solubility, domain boundaries, etc. We offer a variety of expression vectors, including a set of LIC vectors (Ligand Independent Cloning) for protein expression in insect cells and mammalian cells. For expression in insect cells we have SF9 and SF21 systems for expression of intracellular and extracellular proteins. Our mammalian expression system has been optimized for HEK-293T cells, and we also have the glycosylation free, gnT1-deficient HEK293S variety.
Protein expression in bacteria (E. coli) is beyond the scope of the facility!
The NKI Protein Facility is part of the Instruct-NL national Instruct Centre of the Netherlands and provides services for production, purification, biophysical characterisation and crystallisation trials for proteins. Our facility staff is well known for their long-time participation in several EC-funded projects, most recently in iNEXT-Discovery, ISIDORe, canSERV and Fragment-Screen.
The Protein Facility is accessible to internal (NKI) and external research groups, either local, national or from abroad. In particular researchers without extensive experience or facilities to produce and analyse their proteins are invited to apply; they can receive assistance with the design, execution and analyses of their experiments!
Some experiments (e.g. high-throughput macromolecular crystallisation) will be performed by the facility staff. However, for most access possibilities researchers visit us in Amsterdam, not only for expert guidance in on-site experimentation and analysis but also to receive hands-on training. A particular strength of the facility is the characterisation of macromolecular interactions using a broad set of complementary biophysical methods.
To ensure a smooth application process, we do recommend researchers to contact us before preparing and submitting an application.
The expression of soluble, functional proteins depends on many different factors. To increase the chances for successful expression our NKI Protein Facility experts can assist in the design of selected protein fragments using structural and bio-informatics tools.
The NKI Protein Facility houses large-scale incubators at different temperatures for protein expression in E. coli (not offered through Instruct!), and expression platforms for insect cells (using the baculovirus system) and mammalian cells (e.g. a large roller-bottle incubator with a capacity for 90 bottles).
For purification of expressed proteins, a variety of chromatography methods - affinity, ion exchange and size exclusion chromatography - can be requested on our different Akta and BioRad systems.
The NKI Protein Facility combines a multitude of biophysical techniques to characterise (bio)macromolecular properties, stability and interactions. Our equipment includes:
The NKI Protein Facility makes use of an automated high-throughput crystallisation screening platform. There, protein crystallisation conditions can be screened in 96-well format using 100-200 nanoliter droplets. Plates are prepared by (nanoliter) liquid handling robots, and crystallisation trials are monitored over time by automated imaging systems at 4 °C and 20 °C. Crystallisation results are made accessible to our guests via web-servers.
For more information on the availability of the services and machines available at the NKI Protein Facility, please visit the facility website, or contact the facility staff directly.
For overall information on the availability of Instruct-ERIC technologies, visit the Technology Availability List or contact support@instruct-eric.org.
How to find us
The Netherlands Cancer Institute (connected to the Antoni van Leeuwenhoek hospital - NKI/AvL) is conveniently located at the south-west part of Amsterdam. From Schiphol airport it takes roughly 15 minutes to drive to the institute (bus, car/taxi), or 7 minutes by train (to Amsterdam Lelylaan station), followed by a 15 minute walk. From the institute it is easy to connect to Amsterdam city centre by tram, bus or train. More details can be found at the NKI website.
Visit this centre and more by submitting a proposal for access.
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