Protein assay problems
ekhatipo at NOSPAMmidway.uchicago.edu
Wed Jan 24 12:53:24 EST 2001
<engelbert_buxbaum4 at my-deja.com> wrote in message
news:94m3ib$jke$1 at nnrp1.deja.com...
> In article <3A6DBD35.B03B9A51 at hotmail.com>,
> The Grouchybeast <thegrouchybeast at hotmail.com> wrote:
> > I am attempting to do Western blots with material from Laser Capture
> > Microdissection caps.
> > I am reluctant to try to precipitate out proteins before assaying
> > because I have so little protein and I'm worried about creating false
> > variability betwen samples.
> This depends on the method you use. Chloroform/Methanol
> precipitation (Wessel + Fl"ugge) is quite reliable, and if you follow
> that with the BCA protein determination (check the Pierce catalogue) the
> whole thing is quite sensitive (especially in microcuvettes, were you
> can work with 50-100 ul total volume at 10 mm path length)
Could you post the details of the Chloroform/Methanol method? I did not here
about it and would like to try it vs. trichloroacetate.
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