We have a few Strep-tactin columns that had some growth in them and we would like to regenerate the columns back since the resin is quite expensive. Basic goal: remove the brown stuff.

So far, I've killed whatever living material there is with 20% EtOH and 0.5 N NaOH. After cleaning out the columns with Tris Buffer, I've tried regenerating the columns with Buffer with HABA and the resin is a bit redder than before (typical reflection of good regeneration). However, rinsing out the columns again still leaves behind a brownish color. There is also a good deal of black particulate in the resin.

I'm curious what methods exists to clean out a contaminated column.

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    $\begingroup$ I'd check the data sheet of the column for the limits of what the column can handle and just try the harshest of those conditions. They usually also specify cleaning procedures there. I'd try something with high salt to wash out that stuff, maybe also reverse the flow direction and run the column backwards. $\endgroup$ – Mad Scientist Jun 1 '13 at 9:19
  • $\begingroup$ I'm going to try the high salt. That's a good suggestion. The reverse flow probably won't work since I've repacked the column a few times by now $\endgroup$ – bobthejoe Jun 1 '13 at 22:54
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    $\begingroup$ in the future consider azide is also sometimes used to keep columns sterile in storage. Though it won't clear out the dead matter...its possible that the column was never meant to be this cluttered up. In my wet lab days i'd be throwing that out... sorry to be a party pooper, but molecular purity is hard enough without whole cell fragments drifting about. $\endgroup$ – shigeta Jun 2 '13 at 4:02
  • $\begingroup$ @Shigeta, alas it wasn't my column. I guess it's time to buy some new resin. $\endgroup$ – bobthejoe Jun 6 '13 at 19:42
  • $\begingroup$ Yeah sorry, but remember your time is valuable too - don't spend 3 days trying to fix equipment that might ruin your prep...unless you want to be fixing equipment of course. $\endgroup$ – shigeta Jun 6 '13 at 22:37

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