The facility of Protein Technology at CIBIO Department provides expertise in recombinant protein technologies, protein biochemistry and structural biology.
We offer support, training, materials and instrumentation for proteins production, purification and their biophysical characterization.
Services
You can interface with us in many different ways. If you are an experienced protein scientist, you can use the core facility as an extension of your lab, where you will find everything you need to conduct your research. We can also provide training and mentoring to users that are not comfortable with one or more steps of the workflow. In addition to the training and access to the instrumentation available, we can offer different standardized and personalized purification packages. Our main goal is to help the users with the design and implementation of every purification step, to be performed independently or in collaboration with the facility. Full purification services (i.e. carried out entirely from the facility) can be available and will be discussed on a case by case basis, according to the facility capacity, only for well characterized proteins.
The facility has a wide variety of E. coli cell stocks and expression vectors. Small scale test expressions as well as large scale expression can be conducted at different temperatures and conditions, as well as in different enriched media (i.e. for the labeling with 15N, 13C, or selenomethionine). We can provide help in the design and implementation of your expression construct as well as mutagenisis, using traditional cloning and more cutting edge assembly methodology, commonly used in synthetic biology.
Additionally we have expertise in conjugation of purified proteins with fluorophores or other small molecules.
The facility is equipped with FPLCs and a large assortment of chromatography columns. In addition to our available columns, users may use their own column. We have all the materials needed for gravity columns (glassware and different type of resins), dyalisis, protein concentration and quantification.
We also offer our expertise in protein-protein and protein-DNA interactions and we can help you in the design of a variety of fluorescence based assays that you will be able to perform on your own with the facility's instrumentation (i.e. Fluorescence polarization, FRET).
Instrumentation
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AKTA Pure and AKTA Purifier (Cytiva)
These instruments can be used for analytical and large scale purifications using a wide range of chromatographic columns available in the facility or your own column. User can access these FPLCs independently after receiving a training. -
Microcal PEAQ-ITC (Malvern).
Isothermal titration calorimetry can be used to measure dissociation constants, stoichiometry, and thermodynamic parameters of a biomolecular interaction in the nano molar and high micro molar range. -
Dynamic Light Scattering Zetasizer Nano ZS (Malvern).
It can be used to analyse size distribution, molecular weight and zeta potential of protein and nanoparticles. It can also be used as a detector of the new AKTA pure FPLC in tandem with Size exclusion Chromatography. -
Quantamaster Spectrofluorimeter (PTI)
For any fluorescence based analysis of fluorescent molecules, as well as the study of macromolecular interactions through FRET, Fluorescence polarization and kinetic measurements. Our instrument is equipped with a customized stopped-flow accessory for the rapid mixing of samples during kinetic measurements. -
Typhoon 5 (Cytiva) laser scanner
For imaging and precise quantitation of fluorescent, colorimetric and radiolabeled proteins and nucleic acids. -
Thermoshakers are available for large scale expression of proteins from 10 to 37 C.