Protein melt analysis is an inexpensive way to screen many small molecule compounds, aptamers, or other short molecules for their ability to bind to a protein.Protein melt measures an increase in fluorescence when a hydrophobic dye gains access to the inner core of a purified protein target as it denatures. The association of another moiety, if it interacts with a high enough affinity with the target protein, will affect the proteins melt curve through a slight change in protein structure that can be measured. The association of another molecule, if the small molecule interacts specifically with the target protein, that will affect the proteins melt curve through a slight change in protein structure that can be measured. By using a real-time qPCR instrument (Roche LC480), the melt curve can be recorded and analyzed utilizing software for this purpose. Contact the director of the QGCL for more information. Protein concentration would be in the 75-150 µg/ml range, depending on the molecular weight of the protein, and buffer conditions will depend on the conditions that are optimal for the target protein. In any case, the protein will have to be very pure, as with fluorescence polarization studies, to work.
Data analysis- Data will be provided in an Excel workbook. All melt curve figures and sample identification will be provided.