Microsomal Fraction Protocol

Francis Durst francis.durst at bota-ulpnospam.u-strasbg.fr
Sat Mar 25 10:28:17 EST 2000


In article <8beivt$k61$1 at mark.ucdavis.edu> "Alfredo Lopez De leon Ph.D." <allopezdeleon at NOSPAM.ucdavis.edu> writes:
>From: "Alfredo Lopez De leon Ph.D." <allopezdeleon at NOSPAM.ucdavis.edu>
>Subject: Microsomal Fraction Protocol
>Date: Thu, 23 Mar 2000 18:09:04 -0800

>Dear All:
>I am looking for a good protocol to prepare microsomal membranes from rice
>leaves.  I've been trying Yoshida's method (Plant Phys. 1983).  However I
>had to grind the tissue in liquid Nitrogen because it was impossible to get
>anything with a Polytron.  Is there a better method for rice leaves or
>maize, barley, wheat ?  Am I destroying the microsomal fraction by grinding
>the tissue in liquid Nitrogen?

No, you don't. This works fine with the appropriate buffer. There are many 
recepies. Look at papers about isolating fragile microsomal enzymes like the 
P450s. For wheat look at Frear in Pesticide Biochem. Physiol., or Zimmerlin A. 
and Durst F.  Phytochemistry, 29, 1729-1732 (1990). More general 
considerations in Durst et al. (1996)  Methods Enzymol.  272, 259-268

Francis








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