How do we study chaperone interactions?

1. Heat target protein (PrT) at 45C for 15 min and separate aggregated (P, pellet) and soluble (S) fractions by centrifugation. SDS-PAGE, silver stain and quantify. Most is insoluble. 2.Heating in the presence of Hsp40 increases the S fraction. 3. Heating in the presence of Hsc70 has a larger effect 4. Hsp40 and Hsc70 together have even more effect 5. Maximal solubilisation requires Hsp40, Hsc70 and ATP