In <Pine.SOL.3.91.960314134928.3098B-100000 at general4.asu.edu>,
costein at IMAP1.ASU.EDU writes:
>I've been working on ATPase (CF0F1) extraction from spinach leaves, and
>the procedure seems to be straight forward I don't think I have been
>successful in isolating the protein.
>the way I use to test for activity of the protein is by reconstituting it
>with liposomes and shoking the proteoliposomes with an acid and then a
>base and finally adding a luciferin/luciferase reagent.
Are you doing any ATPase measurements as you go along the prepatory
procedure? ATPase can be easily stimulated by methanol, octylglucoside,
and other treatments. The luciferase assay can be challenging enough,
without having to worry whether you have active protein or not.
elarson at ux1.cso.uiuc.edu