Dear Crystallographers, I am interested in doing a type of pull-down experiment by immobilizing protein X on IMAC resin, flowing a large volume of dilute lysate containing protein Y over it, then adding some concentrated agent (solid SDS perhaps) to some more of the same lysate, and running that over the column to elute protein Y off protein X, without eluting X off the column. I am afraid from past experience that SDS might knock X off the column, presumably depending on the concentration. I do not care about the folding state of Y--I will just be running a PAGE gel anyway. Does anyone know either what is the minimal concentration of SDS for robustly unfolding proteins/breaking up interactions (and whether that concentration is safe for IMAC), or what would be a good alternative agent to do the same?
Thanks in advance for your help, Jacob Keller ******************************************* Jacob Pearson Keller Northwestern University Medical Scientist Training Program cel: 773.608.9185 email: [email protected] *******************************************
