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Relative to other affinity interactions, the binding between GST and glutathione is pretty slow, so youll want to incubate your protein of interest with the beads for around 30 minutes to an hour before uncapping the column and letting the flow-through drip out the bottom

Vitamin B12 requires an intrinsic factor-mediated active transport for absorption, therefore, lack of or inhibition of intrinsic factor results in pernicious anemia