A brief guide to why ITC or DSC experiments may not work as expected
Why do ITC experiments not work as expected?
Concentration errors because KD is unknown or different value than expected
- Too little heat change
- Never reach saturation
- Rapidly saturate binding sites
- Incorrect concentrations used
- Buffer mismatch – too large heat of dilution
- Other sources of heat change in system
- No binding
Why do DSC experiments not work as expected?
- Incorrect sample preparation
- Protein already denatured prior to DSC
- Incorrect concentrations used
- Buffer mismatch between reference and sample cell
- Dirty cells
- Incorrect filling technique
- Thermal history not established
What to do next
Once you have determined the cause(s) of the error(s) in your experiment, please contact us for further assistance.