MadSci Network: Biochemistry

Subject: Why there is no lipase activity in my dialysate

Date: Tue May 13 07:01:00 2008
Posted by Sam
Grade level: grad (science) School: No school entered.
City: No city entered. State/Province: No state entered. Country: No country entered.
Area of science: Biochemistry
ID: 1210687260.Bc

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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