Martin Guttenberger, University of Tübingen, Tübingen, Germany
Published online: May 2005
DOI: 10.1038/npg.els.0003916
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
| References | |
| 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. | |
| book 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. | |
| book 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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