determ. of prot. conc. in 8M urea

Robert Kuhelj Robert.Kuhelj at ijs.si
Fri Feb 25 08:10:39 EST 1994


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
denaturant conc.!)?

What happens to the molar absorbance coefficient of the protein 
absorbance of aromatic amino acids increase substantially at elevated
urea conc.!)?

Any experiences?

Robert                               robert.kuhelj at ijs.si




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