Hi,
One of my colleague put the Superdex 200 10/300 GL column mistakenly in 1M NaOH at flowrate of 0.2ml/min overnight instead of water wash. Will the column be stable? How to know that the column is fine and not damaged? Is there any rescue measures that can be taken to nullify or revert the effect of NaOH?
It would be fine. We generally wash superdex 200 with 1 M NaOH, on average in 6 month. After NaOH, wash with water, 2-3 CV and equilibrate with buffer to use it.
Removing severe contamination
1. Reverse the flow.
2. Wash with 4 column volumes of 1 M NaOH (to remove hydrophobic proteins or
lipoproteins) followed by 4 column volumes of distilled water.
3. Wash with 0.5 column volume of 30% isopropanol to remove lipids and very
hydrophobic proteins, followed by 2 column volumes of distilled water.
4. Equilibrate the column with at least 5 column volumes of buffer, or until the baseline
monitored at A280 and the pH of the eluent are stable, before beginning a new separation.
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