Differential Scanning Fluorimetry (a.k.a. Thermofluor) at the C3
Differential Scanning Fluorimetry (DSF) is our primary technique for assessing Protein Stability.
To book a standard DSF test of different buffers at the C3:
- Login to the C3 booking software
- Select the option DSF (under resource bookings) from the sidebar – this will open the calendar view
- Select an empty slot, and click ‘BOOK’
- In the field “Experiment Aim” type “Buffer screen” then fill in the rest of the form as appropriate*
- Then make sure to provide the C3 lab with both the protein sample (>40uL at 1mg/mL or higher) and the base (control) formulation (ideally flow through from purifying/concentrating your protein)
We can also run your protein against the Hampton Solubility and Stability Screen – for this you must provide 50mL of base formulation.
In the same way we optimise for crystallisation, we can optimise for protein stability – the TECAN will allow us to set up specific screen designs. Talk to us if you’re interested in this process.
You can also use DSF to look at comparative stability, by running more than one protein in the same experiment. Commonly, this is used to compare the stability of different mutants of the same protein, or to compare the stabilising effect of different compounds on the same protein. Click here to learn more about how to do these experiments.
When we have run the experiment, we attach the resulting experiment file to the booking form and you’ll receive an email notification.
To read this file, you will need to download the BioRad software (link available on the bottom of every DSF booking form)
READ MORE: Assessing DSF results.