j.reid at sheffield.ac.uk
Thu Oct 26 09:05:13 EST 2000
"overlapping" buffer systems is a very good idea and is easy to do.
Also is is well worth while keeping the ionic strength constant throught
the buffer range. There is an paper in "Methods in Enzymology" that
covers this - I forget the ref. but it might be in vol63, anyway recipes
are also in "The biochemists handbook".
By the way if it is enzyme action that you are intrested in measuring
"activity" vs. pH is not very informative; kcat/Km or kcat vs. pH are
much more intresting (see e.g Knowles 1976 CRC critical reviews in
biochemistry p165, the chapter in the Enzymology labfax, Brocklehurst
1994 Protein Eng. 7,291-299 or any introductory textbook, Fersht has a
good chapter as does Cornish-Bowden.)
All the best,
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