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Urea/Thiourea lysis buffer for Protein Solubilization after Trizol Extraction

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Posted 30 June 2009 - 08:19 AM


There's anyone have any experience with preparation of the following lysis buffer to dissolve protein pellets after Protein extraction with Trizol?

Lysis Buffer:

8M Urea
3M Thiourea
4% CHAPS (0.065 M)
1% DTT (0.064 M)

in water.

I've been struggling to get Urea and Thiourea dissolved in water. It just won't dissolve!
Anyone that has been through the pain, can get me any ideas/ tips?
Do you prepare the solution fresh each time? What volume usually?

Thanks a lot!

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