determ. of prot. conc. in 8M urea
Wakarchu at wakarchu.bio.nrc.ca
Fri Feb 25 17:21:27 EST 1994
In article <1994Feb25.141039.940 at cathy.ijs.si> Robert.Kuhelj at ijs.si (Robert Kuhelj) writes:
>From: Robert.Kuhelj at ijs.si (Robert Kuhelj)
>Subject: determ. of prot. conc. in 8M urea
>Date: 25 Feb 94 14:10:39 +0100
>Any idea how to quickly determine protein concentration in 8M urea solution?
>SDS PAGE is too slow and you can only estimate the amount of protein.
>Bradford or Lowry are probably strongly influenced by such a high urea
>concentration, and the calibration curve must be prepared using the 8M urea
>solutions of the standard.
>WHAT ABOUT SPECTROPHOTOMETRY?
>What wavelength to use (a slight red shift of the spectrum occurs at high
>What happens to the molar absorbance coefficient of the protein
>absorbance of aromatic amino acids increase substantially at elevated
>Robert robert.kuhelj at ijs.si
The classical determination of protein concentrations by A 280 works just
fine denaturants such as urea or guanidine-HCL. There is almost no effect
on the extinction coefficent for trp and tyr (what you see at 280 nm) in
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