As you know, one traditional step after a AS pp is HIC in order to perform a purification removing the high salt concentration at the same time. For this procedure, you usually add AS to just below the concentration necessary to precipitate your protein and you load the supernatant with your target protein in an HIC colunm....but what happen if your protein is able to maintain itself in the supernatant only at very low AS concentration (for example 20% AS), then, probably this low AS concentration is not high enough to bind your protein to HIC colunm and you can not rise the binding buffer salt concentration because your protein can be precipitated in the colunm.
Then, the this system (AS pp + HIC) is not possible for protein which precipitate at very low AS concentrations, am I right?
Thank you for your coments!!!