Hello,
I am looking to extract this protein called methanobactin using chromatography with Diaion-HP20. Its been done before and literature states the following:
This is what the paper says: The filtrate was then loaded onto a 4 × 30 cm Diaion HP-20 column (Supelco). Subsequently, the column was washed with two column volumes of deionized water prior to elution with 60% methanol. The eluate was freeze-dried immediately.
I am only aware of the theory behind chromatography, so there is a lot of information that I feel I am missing.
Elution with 60% methanol. How much should I use? Is this proportional to the amount of HP-20 that I use? How do I find out this?
I am assuming that I also collect all that is filtered since it will be freeze-dried anyways. Is this correct?
What is the solvent in this case? Water?
If someone could explain this in more detail, that would be greatly appreciated.
Adam
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