BIFI-LACRIMA: Sample requirements

Calorimetry: VP-ITC, Auto-iTC200, VP-DSC, and Auto-PEAQ-DSC (MicroCal, Malvern-Panalytical)

What are the sample requirements?

  • Pure samples are required.
  • For a typical DSC assay, you might need 0.5 mL of 20 μM protein solution.
  • For a typical ITC assay, you might need 0.5 mL of 10-20 μM protein solution and 0.15 mL of 100-200 μM of ligand solution.

What other specific considerations are relevant?

  • It is important that protein and ligand solutions are perfectly matched regarding buffer composition.
  • Avoid large concentration of solubilizing agents (e.g., glycerol, DMSO) and reducing agents (e.g., DTT, TCEP).
  • Buffers with low ionization enthalpy are preferred for DSC.
  • Please contact for specific details on the experimental protocol.
  • Contact person: Adrian Velazquez-Campoy ()

Surface plasmon resonance: Biacore T200 (GE Healthcare)

What are the sample requirements?

  • Pure samples are required.
  • The attachment of the macromolecule of study to a functionalized surface (SPR chip) is required.

What other specific considerations are relevant?

  • Please contact for specific details on the experimental protocol.
  • Contact person: Adrian Velazquez-Campoy ()

Microscale thermophoresis: Monolith NT.115Pico (NanoTemper)

What are the sample requirements?

  • Pure samples are required.
  • The attachment of an appropriate fluorophore probe to the macromolecule of study is required.

What other specific considerations are relevant?

  • Please contact for specific details on the experimental protocol.
  • Contact person: Adrian Velazquez-Campoy ()

Spectroscopy: Chirascan spectropolarimeter (Applied Photophysics), Cary Eclipse fluorimeter (Agilent), NanoStar DynaPro DLS and DynaPro Plate Reader III (Wyatt Technology)

What are the sample requirements?

  • Pure samples are required.
  • For a typical assay you might need as much as 0.8 mL of 10 μM protein solution (e.g., near-UV circular dichroism measurement), and as less as 0.05 mL of 5 μM protein solution (e.g., DLS measurement).

What other specific considerations are relevant?

  • Please contact for specific details on the experimental protocol.
  • Contact person: Olga Abian ()

Multimode plate readers: FluoDia T70 (PTI), Synergy HT (BioTek), Mx3005p (Agilent), CLARIOStar and FLUOStar (BMG Labtech)

What are the sample requirements?

  • Pure samples are required (for proteins and chemical compounds).
  • You may need specific cell lines in order to observe cell-specific effects.
  • For a typical assay you might need as less as 0.1 mL of 0.2 μM protein solution (e.g., enzymatic reaction) or as much as 0.1 mL of 2 μM protein solution (e.g., differential scanning fluorimetry).

What other specific considerations are relevant?

  • Please contact for specific details on the experimental protocol and amount of sample required.
  • Contact person: Nunilo Cremades (), Olga Abian ()

Fluorescence microscopy: DMI 6000B (Leica)

What are the sample requirements?

  • Pure samples are required (for proteins and chemical compounds).
  • You may need specific cell lines in order to observe cell-specific effects.
  • Proteins can be introduced into cells, but usually cells are transiently transfected with DNA constructs that code for the desired protein, either alone of fused to fluorescent proteins.
  • Specific stable cell lines expressing the protein of interest can also be generated and analysed.

What other specific considerations are relevant?

  • The attachment of an appropriate fluorophore probe to the macromolecule of study is required for live cell experiments.
  • For immunocytochemistry, specific antibodies are required.
  • Fluorescently labelled ligands can also be used.
  • Please contact for specific details on the experimental protocol.
  • Contact person: Jose Alberto Carrodeguas ()

Single molecule spectroscopy: MicroTime 200 (PicoQuant)

What are the sample requirements?

  • Highly pure samples are required.
  • Standard measurements are performed with 50 μL of 200 pM labelled protein for single-molecule experiments or 1 nM for fluorescence correlation spectroscopy measurements.

What other specific considerations are relevant?

  • The attachment of an appropriate fluorophore probe to the macromolecule of study is required.
  • Please contact for specific details on the experimental protocol
  • Contact person: Nunilo Cremades (ncc@unizar.es)