Hi All,

I recently did the thermal stability shift assay with my 30 kDa monomeric protein with different concentrations. I have noticed that it gives us three Tm as you increase the concentration. How come one monomeric protein gives three Tm at a higher temperature. However, We saw one consistent peak at 70C in all three concentrations of the protein. the additional Tm comes at 90C as we increase the concentration. How would we know the correct Tm from the mixture of three Tm? Should we always select the first peak at the lower concentration of protein? What is the ideal concentration for performing thermal stability shift assay in CFX connect -real time machine?

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