Urea-containing buffers: How long can I store them?
rsteinbe at etowah.uokhsc.edu
Wed Mar 19 20:51:03 EST 1997
Viraj Master wrote:
> Hello all,
> I am using the Qiagen pQE series of vectors to express proteins. There
> are two buffers which I use a lot, Buffer B and Buffer C.
> These contain:
> 8M urea
> 0.1M NaH2PO4
> 0.01M Tris
> Buffer B is at pH=8
> Buffer C is at pH=6.3
> I have been told that I must make these buffers up **fresh** each time
> prior to use. I would like to be able to prepare batches of these
> buffers and store them. Does anybody know whether this can be done and
> not affect the protein prep? If so, are they stored in the cold, or at
> room temp? For how long can they be stored?
> Many thanks,
> Viraj Master PhD
> Dept of Organismal Biology
> University of Chicago
I assume the the problem is the urea, which can decompose forming
isocyanate-- this, in turn, will react with your proteins. Although I
would not recommend storing urea-containing buffers for extended
periods, you can alleviate the isocyanate problem by including a high
concentration of a scavenger (e.g., L-lysine) in your buffers.
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