Protein Determination by UV Absorption

  • Alastair Aitken
  • Michèle P. Learmonth

Abstract

Quantitation of the amount of protein in a solution is possible in a simple spectrometer. Absorption of radiation in the near UV by proteins depends on the Tyr and Trp content (and to a very small extent on the amount of Phe and disulfide bonds). Therefore the A280 varies greatly between different proteins (for a 1 mg/mL solution, from 0 up to 4 [for some tyrosine-rich wool proteins], although most values are in the range 0.5-1.5 [1]). The advantages of this method are that it is simple, and the sample is recoverable. The method has some disadvantages, including interference from other chromophores, and the specific absorption value for a given protein must be determined. The extinction of nucleic acid in the 280-nm region may be as much as 10 times that of protein at their same wavelength, and hence, a few percent of nucleic acid can greatly influence the absorption.

Keywords

Quartz Filtration Albumin Citrate Sodium Chloride 

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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Alastair Aitken
    • 1
  • Michèle P. Learmonth
    • 2
  1. 1.Division of Biomedical and Clinical Laboratory Sciences, Membrane Biology GroupUniversity of EdinburghScotland, UK
  2. 2.Department of Biomedical SciencesUniversity of EdinburghScotland

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