Surface Plasmon Resonance – Biacore T200

What is it used for?

Real-time detection and monitoring of biomolecular binding events

  • Specificity – How specific is the binding between two molecules
  • Concentration – How much of a given molecule is present or active
  • Kinetics – What is the rate of association / dissociation
  • Affinity – How strong is the binding?

Interactions between proteins, nucleic acids, lipids, carbohydrates and even whole cells can be studied (molecules as small as 100 Da).

Key features

  • Sampling from tubes, 96 or 384 well plates
  • 4 flow cells
  • Determination of fM to mM dissociation constants (Kd), with sample volumes from 50 to 400 ul
  • Concentration range 1 pM to 2 mM
  • Temperature control from 4 to 45°C

Examples of how the WISB community are using this equipment

  • Determination of binding constants for a selection of DNA-binding proteins (PI Corre)
  • Determining the affinity of protein-protein and protein-ligand interactions (PI Challis)

MicroScale Thermophoresis – Monolith NT.115

What is it used for?

Measurement of biomolecular interactions – from the formation of protein complexes to the binding of single metal ions.

Key features

  • Nano Blue/Red detector channels for fluorescence
  • Determination of nM to mM dissociation constants (Kd), using very small amounts of sample (4 ul)
  • Required concentration of fluorescent molecule 10-9 – 10-3 M
  • Temperature control from 22 – 45°C

Examples of how the WISB community are using this equipment

  • Determination of substrate specificity and binding constants for nutrient uptake transporters from Mycobacterium tuberculosis (PI Fullam)
  • Identification of protein-protein interactions between eIF4E4 and PABP1, proteins that are part of Leishmania’s translation initiation complex (PI McCarthy)

WISB has been funded by the above bodies