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<DIV>Hi,</DIV>
<DIV>I am trying to extract Rubisco from grapvine leaves, and I generally end up
with none. I use standard extraction buffers (200 mM Tris.Cl pH8.0, MgCl2, EDTA,
etc.) and I have to use up to 10% insoluble PVPP and up to 5% Triton X100 to
actually see some rubisco in SDS-PAGE gels. I believe that the problem is
that Rubisco is binding to phenols and its is being pulled down during the
centrifugation step. Triton does help to solubilize some of this aggregates,
however I do not get consistent results (sometimes works but most of the time it
does not...). I believe it must have to do with plant's growing conditions (some
leaves are more phenols than others). As anybody run into a similar problem? Any
sugestions on how to solubilize Rubisco (in active form...) from whatever
agregates its doing? Thanks a lot.</DIV>
<DIV>Joao </DIV>
<DIV>**********************************************<BR>Joćo P. Maroco<BR>Lab.
Ecofisiologia Molecular - ITQB<BR>Av.Republica. EAN<BR>2780
Oeiras<BR>Portugal<BR>Phone: 214469641<BR>Mobile:965447096<BR>Fax:
214433644<BR>e-mail: <A
href="mailto:jpmaroco@itqb.unl.pt">jpmaroco@itqb.unl.pt</A></DIV></BODY></HTML>
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