Measuring Protein Concentration with Absorbance, Lowry, Bradford Coomassie Blue, or the Smith Bicinchoninic Acid Assay Before Electrophoresis

Methods Mol Biol. 2019:1855:31-39. doi: 10.1007/978-1-4939-8793-1_3.

Abstract

Measuring the concentration of proteins is an essential part of enzyme analysis or serves to monitor protein yields and losses during protein isolation procedures. Decisions on the usefulness of any protein isolation procedure depend on knowing the concentration of proteins before and after a procedure. Protein concentration in solution is generally measured with spectrophotometry in the UV range or in the presence of dyes or copper interacting with the protein. This review describes absorbance at 280 nm, the Lowry, Bradford (Coomassie Blue), and Smith (bicinchoninic acid) assays for measuring protein and includes suggestions for optimizing each method.

Keywords: Absorbance; Bicinchoninic acid; Bradford; Coomassie; Lowry; Protein concentration; Protein staining.

MeSH terms

  • Indicators and Reagents / chemistry*
  • Proteins / analysis*
  • Quinolines / chemistry
  • Rosaniline Dyes / chemistry
  • Spectrophotometry, Ultraviolet

Substances

  • Indicators and Reagents
  • Proteins
  • Quinolines
  • Rosaniline Dyes
  • bicinchoninic acid
  • coomassie Brilliant Blue