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Protein Production

  • Provision of a guided work flow
  • Microbial, insect and mammalian cell hosts
  • Cell free in vitro expression in various host lysates

Protein Production Core Facility

The Protein Production Core Facility (PPCF) is designed to provide users with the complete infrastructure for protein production using the most modern and efficient methods. Particular attention is paid to maintaining the highest quality standards of the samples to be produced, which guarantees the suitability of the samples for functional and structural studies in a reproducible manner. Additionally, we aim to develop sustainable processes.

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Services

The Protein Production Core Facility offers all steps of protein production consisting of the mutagenesis and cloning of target genes in expression vectors, their heterologous expression in prokaryotic and eukaryotic hosts, as well as the subsequent purification of the corresponding proteins as a service. In addition to the full service, in which the employees of the facility carry out the work, the users can carry out the work in the facility under the supervision of the facility team. The service includes advice on the selection of suitable strategies, the provision and optimization of suitable protocols, the instruction and use of the required equipment and the provision of the required vectors, cell lines, chromatography materials and all necessary consumables. The following services are offered in detail:

- Cloning and mutagenesis
- Cultivation of prokaryotic and eukaryotic cells of risk group R2 or security level S2
- Transformation of prokaryotic cells
- Transfection of insect cells and mammalian cells
- High-throughput heterologous expression screening in plate format: E. coli & insect cells
- Heterologous expression in E. coli, insect cells and mammalian cells
- Cell harvest and cell lysis
- Ultracentrifugation and cell membrane isolation
- Protein purification using Äkta Pure chromatography devices
- Development and optimization of protein purification protocols
- Automation of protein purification protocols
- Biochemical characterization (SDS and native PAGE, IEF, Western Blotting)
- Biophysical characterization (stability measurements using microscale thermophoresis)
- E. coli strain collection
- Expression vector collection
- NEB freezer

Available Instruments

  • Equipment for cloning and mutagenesis projects
  • PerfectBlue Gelsystem Mini S, M, L
  • Eppendorf Mastercycler X50a – PCR Cycler
  • Bio-Rad ChemiDoc MP – imaging system
  • Bio-Rad  MicroPulser – electroporator for transformation of bacteria
  • Infors HT Minitron – incubator for insect cell expression
  • Eppendorf  Innoa S44i – biological shaker for bacterial cell culture
  • Heracell Vios 250i – CO2 incubator for adherent cells (Please delete this sentence )
  • Eppendorf  Innoa S41i – biological CO2 shaker for mammalian cell culture
  • Bandelin Sonoplus - sonification equipment
  • Various Eppendorf Centrifuges 5920R - multi use centrifuge
  • Eppendorf Centrifuge 5430R - refrigerated centrifuge
  • Various Eppendorf Thermomixer C/F1.5
  • 3x Thermo Fischer MaxiSafe 2020 – laminar flows
  • 2x Beckman Coulter Avanti JXN-26 – harvesting centrifuges
  • Beckman Coulter Optima Max-XP and Optima XPN-90 – ultracentrifuges
  • GE Healthcare Äkta pure - protein chromatography system
  • Various BioRad power pacs (basic and HC)
  • XCell SureLock Mini-Cell - PAGE system for self-made and ready-made gels
  • BioRad trans-Blot-Turbo - semi dry Western blotter
  • Tecan SPARK – plate reader
  • Thermo Fischer Micromodulyo – freeze dryer
  • Büchi Rotavapor R-114 – rotary evaporator (Please delete this sentence )
  • Leica Fluorescence cell culture microscope
  • Olympus CKX31SF – inverted cell culture microscope
  • Eppendorf Cryocube F101h - deep temperature freezer

Further information

Access to the PP services and resources

  • The facility is a fee-for-service (non-profit) technology platform operated by UKE. We provide support and expertise for local scientists, the international research community and users from industry.
  • All users must follow a safety training before working in our laboratories. All users are required to undergo training before access to the individual instruments is granted. Currently the biosafety training is given onsite.
  • Please get in touch with us to discuss your specific needs.

 

Our acknowledgement policy

Please acknowledge any substantial contributions of the PP Facility to your research in publications. Please state the following in the acknowledgement section of your publication: “We acknowledge technical support by the PP facility at CSSB Hamburg”.

Please don't forget to let us know when you publish papers that acknowledge us. This will help us keep track and to justify the existence of the facility to funding bodies.

 

PP Facility

Contact

2

Facility Head

Dr. Susanne Witt
Phone:+49 040 8998 87567

E-Mail

Philipp Lewe
Phone:+49 40 8998 87569

E-Mail

Location

CSSB Centre for Structural Systems Biology
c/o Deutsches Elektronen-Synchrotron DESY
Notkestraße 85, Building 15
D-22607 Hamburg

FACILITY OPERATOR

University Medical Center Hamburg-Eppendorf (UKE)

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