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Updated: Jun 12, 2026

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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The lowry method for protein quantitation.

J H Waterborg1, H R Matthews

  • 1Department of Biological Chemistry, School of Medicine, University of California, Davis, California.

Methods in Molecular Biology (Clifton, N.J.)
|June 1, 2010
PubMed
Summary
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Accurate protein concentration determination is best achieved through acid hydrolysis and amino acid analysis. However, the widely used Lowry method offers acceptable protein estimations for complex samples like protein mixtures and crude extracts.

Area of Science:

  • Biochemistry
  • Analytical Chemistry

Background:

  • Accurate protein concentration is crucial for biological and chemical research.
  • Many protein quantification methods are influenced by amino acid composition, limiting absolute accuracy.
  • The Lowry method is a widely adopted technique for protein estimation.

Purpose of the Study:

  • To evaluate the accuracy and applicability of different protein determination methods.
  • To compare acid hydrolysis/amino acid analysis with the Lowry method for protein quantification.
  • To establish the reliability of the Lowry method for practical applications.

Main Methods:

  • Acid hydrolysis followed by amino acid analysis for absolute protein determination.
  • The Lowry protein assay for comparative analysis.

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  • Evaluation of method sensitivity and dependence on amino acid composition.

    • Main Results:

    • Acid hydrolysis and amino acid analysis provide the most accurate protein concentration.
    • The Lowry method's sensitivity is moderately consistent across different proteins.
    • Lowry protein estimations are a practical alternative to absolute determination for protein mixtures and crude extracts.

    Conclusions:

    • While not providing absolute values, the Lowry method is a reliable and acceptable alternative for protein quantification in many research scenarios.
    • The consistency of the Lowry assay makes it suitable for comparative studies and routine analysis.
    • Researchers can confidently use Lowry protein estimations for crude extracts and protein mixtures.