I've been working on ATPase (CF0F1) extraction from spinach leaves, and
although
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. As I haven't
been able to observe any luminescence, I tend to think that I failed to
isolate the ATPase or that it was somehow denatured during the isolation
or the purification procedure.
The gels suggest I have my protein and an additional band at 29000,I am
not sure if that has any significance. In addition although most papers
suggest storing the protein at -180C I have been storing it (for a week)
at -80C, again I am not sure if that has any significance.
If anyone who has isolated CFoF1 has any suggestions or tips I
would be very glad to have them.
my e-mail is costein at imap1.asu.edu
