Can anyone advise me regarding the manual packing/ preparation of gel filtration column using Dextran resin G-75. Details about the column, methods and materials?
The attached booklet entitled "Gel filtration Principles and Methods " describes all information needed for a manual preparation of gel filtration column.
Dear Timir Sir, Thank you very much for your valuable comments, giving details about the column preparation. I would also like to know few more things. Like in ACTA we have detecter so, we know the elution peak for the target protein. How about in this manual method where we don't use detecter? How to collect the fraction of proteins and how to decide which is our target protein? if proteins are similar in molecular weight then? Kindly guide me about this.
u have to elute the fractions in say 1-2 mL volume (depending on your column length) then take reading at 280 nm manually for each fraction. plot a curve and u will get peaks, pool of the fractions of your desired peak. run sds.