What glycerol concentration is appropriate for storage of membrane protein in -80? It seems that 10%-30% is causing aggregation after I store it for 10 days.
Try not using glycerol but low concentrations of detergent instead. See my answer to the previous related question on aggregation of membrane proteins.
You can use 25 - 50 % of glycerol safely in any buffer of your choice (e.g., Tris/HCl or any other). In typical protein storage buffer (evident from published literature), you may consider adding EDTA (I-5mM), DDT (1-5mM), BSA (10mg/ml), protease inhibitors and sodium azide (0.02-0.05%).
Hi Joe,
I completely agree with Jose. Try storing your protein in low concentration of detergent solution. You can read further other RG thread where similar question has been answered extensively. I and Jose also contributed there.
https://www.researchgate.net/post/What_is_the_best_way_to_prevent_membrane_protein_aggregation?ev=tp_feed_post_xview
I have found for membrane preps that sucrose works well. The concentration is 0.25M, but this was for membranes that were not solubilized. As a suspension, it lasted forever... Well at least a year or more. We did this for TNF alpha converting enzyme, a disintegrin and metalloproteinase, a single spanning membrane protein.
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