Hello,
I would appreciate any input from protein experts on how to stabilize a
purified
protein (obtained by overexpression in E.coli).
This particular enzyme is purified using Ni-NTA resin, being eluted with
20 mM
imidazole, 300 mM NaCl and 50 mM NaH2PO4. The enzyme is fully functional
immediately after purification. However, it is very unstable. Storing,
even for a
few hours at 0-10 C (ice or fridges) knocks-out most of the activity.
Same for
-20C. Addition of various %s of glycerol did not help at all. I would
greatly
appreciate any suggestions of additives that could be tried to stabilize
the
enzyme.
Any suggestions for additives that may also stabilize it during
functional
assays are welcome. I realize that this is potentially a more tricky
challenge
(the colorimetric assay is limited in terms of pH and salts etc.).
Stabilizing it
during storage would be a great step.
Thank you,
Pedro Rocha
