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Dear All,
We are esimating protein in S9 fraction in rat liver by lowry method.
After precipitating S9 with 5 % TCA, then adding 0.1 N potassium
acetate ,etanol, isopropyle ether, then centrifuge for 2min at 3500
rpm. I'm getting 2 layers the lower is clear one and the upper layer
precipitated . My doubt is which portion should we taken to estimate
protein in S9 .And what is the normal concentration of protein in rat
(Wisterand SD) S9 fraction , Microsomes.
Please try to help in this regard.
saradhi.
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In our lab after the three wash stages with acetone etc we used to
dissolve the pellet in conc. NaOH for several hours in a heating block
(some people would even leave it overnight at 60C).... microsomes are
not very soluble as you know so dissolving in NaOH would bring them
back into solution.
I do not know if this would be of any help...
Julia Zayezdnaya Zack, Pharm.D.
juliazz.aaa.yahoo.com
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