cleaning chelating sepharose column

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mats978's picture
cleaning chelating sepharose column

Hello, I was hoping someone would be able to help me figure out how to clean cellular extract out of a chelating sepharose column. After bursting cells open I had forgotten to spin down the cellular debris before running it through the column (no sleep and lack of coffee doh). Needless to say the column is caked with junk.
Is there anyway to clean it or should I just scrap it? So far I've tried 70% ethanol and water. The cobalt that was originally on there was stripped with EDTA.
Any help would be greatly appreciated, thanks!

kjosephs's picture
If the collumn has filter

If the collumn has filter frits that you can change then changing or cleaning those will help.  You can also try reversing the flow and runing 0.1N HCl and then 0.1 N NaOH. You might have to junk it though.