Posted 02 July 2013 - 10:55 AM
I have a chromatography issue and could use some advice. I am currently trying to purify a 196 KDa DNA binding protein. The protein binds well to either heparin or DEAE. The problem is getting it off the resin. More specifically getting it off all at one elution step. I am loading the columns at 50 mM NaCl and using a step gradient to elute. The protein begins to come off at around 200 mM. But then it won't even near all elut at that salt concentration. Most of it stays bound and then more will come off at 300mM, then more at 400mM and so on until it pretty much all finally gets bumped off somewhere over 500mM.
Any thoughts would be appreciated
Posted 03 July 2013 - 04:15 AM
if that's not pure enough then you could use a gradient and only pool the fractions that meet your purity requirements.
genius does what it must
i do what i get paid to do
Posted 07 July 2013 - 02:45 PM
Edited by labtastic, 07 July 2013 - 02:47 PM.