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
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Jacob Pearson Keller
Northwestern University
Medical Scientist Training Program
cel: 773.608.9185
email: [log in to unmask]
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