Protein Facility Support and Services
The NKI Protein Facility is
open available for Academic scientists. Both NKI- as
well as external scientists can apply for projects.
A list of support and services is given
below. For more details please check the individual
pages. If your request is not available from the list,
please contact us to discuss the possibilities.
Initial experiments can be
performed by the Facility members, however the aim of the facility
is to assist researchers with performing their own
experiments. For each individual project it will be discussed up to
what level the Facility will execute the experiments and
when the applicant has to provide hands-on support. A
number of experiments will be provided by the Facility
as a service, e.g macromolecular crystallisation.
Construct Design using
CCD (Crystal Construct Designer)
web application for selecting optimal constructs.
Recommendation for cloning,
including assistance with LIC (Ligation Independent Cloning).
A series of LIC vectors is available
from the Facility
Protein expression tests
in E. coli T1 Phage-resistant cells at small scale.
Large scale protein expression in E.
coli T1 Phage-resistant cells, up to 6 liters per batch.
Production of Baculovirus viral
Expression of proteins from
baculovirus in insect cells, up to 4 liters per batch.
Mammalian expression system, using
HEK293 and derivatives like Glycosylation deficient HEK293
strain (GnT-) and HEK293 Flp-In cells for creating stable cell
purifications with affinity resin (Ni-NTA-,
Glutathion beads etc.).
Äkta FPLC systems are used for
protein purification by automated column chromatograpgy.
A variety of columns are available,
including Anion-, Cation-, Size Exclusion (Gelfiltation)- and
calorimetry (ITC) for studying macromolecular
interactions, including protein-protein,
protein-small molecule, protein-DNA and
Surface plasmon resonance (SPR,
BIACORE) for analysis of interaction between protein-protein,
Fluorescence Polarisation for
binding analysis of fluorescent-labeled molecules to proteins
Analysis of protein thermal
stability using thermofluor assay.
of protein multimerisation / complex formation via molecular
mass determination using multi angle laser light
scattering (MALLS) coupled to size exclusion chromatography
screening of crystallization conditions in 96-well
format (vapour diffusion) in nanoliter volumes.
crystallization conditions using TOPAZ microfluidics
system (96-well format).
Optimization of crystallization