protein expression and purification facility (pepf)

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Services offered to members of the Helmholtz Zentrum München

Cloning

The PEPF supports the following cloning technologies

  • Cloning using restriction enzymes
  • Gateway cloning technology
  • Ligase independent cloning (LIC)
  • Sequence and ligase independent cloning (SLIC)

For all these cloning technologies appropriate vectors are available (see Materials).

 

Small-scale bacterial expression screening

For the small-scale expression screening in Escherichia coli we have developed the so-called 'Fast Soluble Expression Screen'.

 

Large-scale bacterial protein production

Target proteins will be expressed on a large-scale (1-10L) in the best E. coli expression host and under optimal expression conditions. The expressed proteins will be subsequently purified using affinity, ion exchange and/or size exclusion chromatography.

 

Protein characterization

Protein characterization of targets proteins using the following techniques:

  • Dynamic Light Scattering (DLS) to determine the size and the monodispersity of a target protein.
  • Static Light Scattering (SLS) to determine the oligomeric state of a target protein and the stoichiometry of protein-protein and protein-ligand complexes.
  • Isothermal Titration Calorimetry to study ligand binding.
  • Thermal shift (ThermoFluor) Assay to determine protein stabiliy and study ligand binding and protein function.