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Related Experiment Video

Updated: Jun 19, 2026

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
06:09

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Quantitation of protein.

James E Noble1, Marc J A Bailey

  • 1National Physical Laboratory, Teddington, Middlesex, United Kingdom.

Methods in Enzymology
|November 7, 2009
PubMed
Summary
This summary is machine-generated.

Accurately measuring protein concentration is crucial in biochemistry. This study reviews various spectrophotometric and dye-based assays, highlighting their limitations and interference issues for selecting the best method.

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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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Last Updated: Jun 19, 2026

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
06:09

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
10:37

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Published on: November 15, 2017

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
09:04

Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification

Published on: August 17, 2015

Area of Science:

  • Biochemistry
  • Analytical Chemistry

Background:

  • Protein concentration measurement is vital for biochemical research, enzymatic studies, and biopharmaceutical quality control.
  • Spectrophotometric assays, utilizing UV-Vis spectroscopy, are common for rapid protein quantitation relative to standards or extinction coefficients.

Purpose of the Study:

  • To review and compare various protein quantitation assays.
  • To inform researchers on assay selection by discussing limitations and interferences.

Main Methods:

  • UV protein spectroscopy
  • Dye-based absorbance assays (BCA, Lowry, Bradford)
  • Fluorescent dye-based assays
  • Amine derivatization assays
  • Detergent partition assays

Main Results:

  • No single protein assay is universally dominant due to method-specific limitations.
  • Dye-based assays can be susceptible to interference from common biological buffer components.
  • Guidance is provided on assay selection considering potential interferences.

Conclusions:

  • Careful consideration of assay limitations and potential interferences is essential for accurate protein quantitation.
  • Selecting the appropriate assay depends on the specific sample matrix and experimental context.