Hi all,
I am currently trying to purify an IgG from Ascites that I have already pulled down on a Protein Agarose A/G bead column. Essentially I am trying to remove as much salt as possible using a buffer exchange and reconstituting in PBS. I currently have 3 options available to me:
Is there any benefit to using one of these over the others? I have mostly been using the Amicon Ultracentrifuge Filters, but even after 2 filtrations, the samples were run on a gel to find the proper isoelectric point and did not run properly, making me suspect it was possible that they still had some salts.
Any help would be appreciated.
Kutibh, Here is a comprehensive discussion on different buffer exchange method: https://www.researchgate.net/post/What_is_the_benefit_of_desalting_by_dialysis_or_spin_columns_in_the_protein_purification_process
It really depends on your prioritized needs: yield, time, and convenience, etc. In addition, if you are going to further purify your sample using IEX, you can choose to use some salt tolerant IEX products to reduce your buffer exchange/dilution burden.
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