Service/Technology

The INSPIRED-A.U.A is part of the Instruct-HL National Instruct Centre in Greece and provides support for the production, purification, biophysical characterization and crystallization of proteins and small molecules. It is accessible to both internal (Instruct-HL) and external research groups. In particular researchers who don't have the experience or the facilities to produce and analyze their specimens can request for assistance. Some experiments (e.g. high throughput macromolecular crystallisation) will be performed by the facility members and are offered as a service. A particular strength of the facility is in the characterisation of macromolecular interaction with a multitude of biophysical methods. Importantly, to ensure a smooth application process, we strongly recommend any users to contact us before submitting an application.

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+ Construct design and cloning

- Expression of soluble,functional proteins is dependent on many different factors. The facility can assist in the design of selective protein fragments using structural- and bio-informatics tools to increase the chance for success.

+ Protein expression in E. coli

+ Purification

ACTA system

+ Biophysical & Biochemical characterization

The facility offers a variety of biophysical techniques to characterise both macromolecular and small molecules' properties and interactions. These include:
- UV-Vis spectophotometer
- Nanodrop
- SDS-PAGE electrophoresis

+ Crystallisation screening

The facility has a variety of crystallisation screening Kits and incubators at 4 °C and 18 °C. Crystallisation conditions can be screened either in 96-well plates or in 24-well plates in milliliter droplets manually. Crystallisation trials are are monitored and visualized over time. Images of high quality can be obtained.

+ X-ray Crystallography

The D8 Venture system includes:
- IμS microfocus X-ray source, Cu radiation, air cooled
- Four-circle goniometer, computer-controlled, crystal-detector distance = 35-240mm, sphere of confusion of less than 7 micrometer
- Detector PHOTON 100, CMOS technology, air-cooled, 100 x 100mm active area, 240e/Cu sensitivity, shutterless data collection
- 800 Series Cryostream cooler, Oxford Cryosystems, AD51 Dry Air Unit, Integrated Autofill system, 60 lt and 150 lt dewar vessels.
- APEX3 and PROTEUM software