Protein Determination


Protein determination aims at estimating the concentration of protein by comparing the unknown sample with a known standard. Sample preparation and choice of a suitable standard are the most important criteria for the accuracy of the results.

Keywords: Bradford assay; dot‐blot assay; interference; Lowry assay; protein determination


Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry 72: 248–254.

Colas des Francs C, Thiellement H and De Vienne D (1985) Analysis of leaf proteins by two‐dimensional gel electrophoresis. Plant Physiology 78: 178–182.

Gallagher SR, Carroll EJ Jr and Leonard RT (1986) A sensitive diffusion plate assay for screening inhibitors of protease activity in plant cell fractions. Plant Physiology 81: 869–874.

Guttenberger M (2004) Protein determination. In: Celis JE (ed.) Cell Biology. A Laboratory Handbook, San Diego: Academic Press (in press).

Guttenberger M, Neuhoff V and Hampp R (1991) A dot‐blot assay for quantitation of nanogram amounts of protein in the presence of carrier ampholytes and other possibly interfering substances. Analytical Biochemistry 196: 99–103.

Guttenberger M, Schaeffer C and Hampp R (1994) Kinetic and electrophoretic characterization of NADP dependent dehydrogenases from root tissues of Norway spruce (Picea abies [L.] Karst.) employing a rapid one‐step extraction procedure. Trees 8: 191–197.

Hoffmann EM, Muetzel S and Becker K (2002) A modified dot‐blot method of protein determination applied in the tannin–protein precipitation assay to facilitate the evaluation of tannin activity in animal feeds. British Journal of Nutrition 87: 421–426.

Lowry OH, Rosebrough NJ, Farr AL and Randall RJ (1951) Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193: 265–275.

Peterson GL (1979) Review of the Folin phenol protein quantitation method of Lowry, Rosebrough, Farr and Randall. Analytical Biochemistry 100: 201–220.

Sokal RR and Rohlf FJ (1994) Biometry: The Principles and Practice of Statistics in Biological Research. New York: Freeman.

Ünlü M, Morgan ME and Minden JS (1997) Difference gel electrophoresis: a single gel method for detecting changes in protein extracts. Electrophoresis 18: 2071–2077.

Zor T and Selinger Z (1996) Linearization of the Bradford protein assay increases its sensitivity: theoretical and experimental studies. Analytical Biochemistry 236: 302–308.

Further Reading

Hjelmeland LM and Chrambach A (1984) Solubilization of functional membrane proteins. Methods in Enzymology 104: 305–318.

Kirazov LP, Venkov LG and Kirazov EP (1993) Comparison of the Lowry and the Bradford protein assays as applied for protein estimation of membrane‐containing fractions. Analytical Biochemistry 208: 44–48.

Marshall T and Williams KM (1992) Coomassie blue protein dye‐binding assays measure formation of an insoluble protein–dye complex. Analytical Biochemistry 204: 107–109.

Neuhoff V, Philipp K, Zimmer H‐G and Mesecke S (1979) A simple, versatile, sensitive and volume‐independent method for quantitative protein determination which is independent of other external influences. Hoppe‐Seyler's Zeitschrift für Physiologische Chemie 360: 1657–1670.

Neuhoff V, Ewers E and Huether G (1981) Spot analysis for glycoprotein determination in the nanogram range. Hoppe‐Seyler's Zeitschrift für Physiologischen Chemie 362: 1427: 1434.

Shakir FK, Audilet D, Drake AJ III and Shakir KMM (1994) A rapid protein determination by modification of the Lowry procedure. Analytical Biochemistry 216: 232: 233.

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Guttenberger, Martin(May 2005) Protein Determination. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0003916]