|MadSci Network: Biochemistry|
I am doing protein purification of lipase from fish. After I did precipitation and recover the pellet, I did a dialysis for this pellet suspension for 12 hours. However, I was not able to find any activity in the dialysate. When I load my dialysate onto an affinity column, I was able to find very little enzyme activity. Why can't I find any activity in my dialysate while there is little activity in my affinity step? How could I improve the dialysate condition so that I could find some activity in the dialysate and obtain more activity in my affinity step? I found out that the protein is not very stable, and dialysis and affinity step were performed at 4 degree in a fridge. Thank you!
Re: Why there is no lipase activity in my dialysate
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