Why does the column turn brown?
The column will only turn brown if there is a reducing agent such as DTT or beta-mercaptoethanol (BME) in the buffer(s). The reducing agent complexes with the cobalt ion to form a brown precipitate. Using up to 10 mM BME should not affect the performance of the resin significantly. If BME is not used, the resin has turned brown, and there is no protein binding, DTT or DTE may be present in the buffers.
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