The manual for Sephadex LH-20 says this: "Resuspend and pour the media slurry into the column in one continuous motion. Fill the reservoir to the top with buffer."
However, I saw another procedure which says: "Fill the column reservoir to the top with solvent."
Does packing this include buffer or just solvent?
Jack Silver
It depends on your purification, and the sort of compound you are purifying.
LH-20 is often used with no buffer, especially when the purification uses only organic solvents. some aqueous purifications don't need buffer, either, while some do.
It is common to buffer proteins; some smaller molecules with ionizable functional groups also benefit from buffering.
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