Protocols for Biophysical Techniques
This page presents a series of short protocols for different biophysical techniques and common aspects of sample preparation. All of the protocols are prepared by the staff of the Biophysics Facility, though some draw on material from other sources. Please do contact us if you would like further explanation or clarification.
- Protein Concentration: A protocol for measuring the concentration of proteins by UV absorbance.
- Freezing and Storing Proteins: A protocol for freezing and storing protein solutions to avoid loss of activity and to achieve consistency in assays.
- Preventing Protein Aggregation: A protocol for the detection of protein aggregates by biophysical methods, and a list of strategies to prevent aggregation.
- Denaturation Fit: A protocol for fitting thermal and chemical denaturation data for proteins using data from any non-calorimetric method.
- Binding Fit: A protocol for fitting 1:1 binding data from any non-calorimetric method, using raw data and accounting for ligand-depletion effects.
- Protein-Ligand Titrations: A protocol for preparing a dilution series for a protein-ligand binding titration.
- CD Thermal Melt: A protocol for acquiring Circular Dichroism thermal denaturation data using a continuous temperature-ramp on the Chirascan Spectropolarimeter.
- CD Unit Conversion: A protocol for converting Circular Dichroism spectra from machine units into standard units for the fitting of secondary structure content and comparison of data from different proteins.
- ITC with Co-solvents: A protocol for performing Isothermal Titration Calorimetry experiments in the presence of co-solvents such as DMSO.