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 Dialysis after Trysol for Westerns [View Printable]
max_rempel

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Joined: Aug 29, 2007







 Send a personal messsage to max_rempel Reply with a quote from this post Go to the top of the page

The Trizol protocol suggests for extractrion of total protein: Dialyze the phenol-ethanol supernate against three changes of 0.1% SDS at 2 to 8°C. Centrifuge the dialyzed material at 10,000 x g for 10 minutes. Use the clear supernate for Western blotting.

I have valuable mouse epidermis samples collected in Trizol (50 mg tissue in 1 ml Trizol) which I will use for RNA for Taqman assay. I wonder if I should try dialysis as described above for Protein analysis by Westerns and Elisa.

Specifically, I am worried that Google doesn't show anyone who does dialysis of proteins after Trizol. It seems to me that the method either doesn't work or is not reliable (which is the same).

If dialysis of Trizol works, I would like to know which membranes to use? Would cellulose membrane be affected by phenol?

Thanks!
.........................

Posted Aug 29, 2007, 20:23 PM
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