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Introduction to the Colorimetric Assays for Proteins

If you are looking for a brief refresher on the Bradford assay and would rather skip the catalogued summary of various colorimetric assays, review it in the designated section.

Brief summary table of colorimetric assays for protein determination
Colorimetric Assays ›››
 Amido Black Method Bicinchoninic Acid Biuret Bradford Assay Colloidal Gold Modified Lowry
Reference Schaffner W, Weissmann C Smith PK Lubran MM Bradford MM Rohringer R, Holden DW Lowry OH
Range [µg/ml] 2 to 24 20 to 200 1,000 to 10,000 1 to 20
(micro);
100 to 1,500
(macro)		 0.02 to 0.64 2 to 100
Volume [ml] 2.0 1.0
("upscalable" for larger cuvettes)		 5.0
("downscalable" for smaller cuvettes)		 1.0
(micro);
5.5
(macro)		 1.0
("upscalable" for larger cuvettes)		 1.0
("upscalable" for larger cuvettes)
Standard Spectrophotomer Wavelength 630nm 562nm 550nm 595nm 600nm-750nm
Preferential Dye-binding Cu+2 interaction with protein basic and aromatic amino acid residues Cu+2 catalyzed oxidation of aromatic acids
Accuracy +++ +++ +++ +++ ++ +++
Convenience + +++ +++ ++++ + ++
Major Interfering Agents none ammonium sulfate,
lipids,
strong acids		 ammonium salts basic buffer conditions,
detergents,
flavonoids		 strong bases ammonium ions,
carbohydrates,
detergents,
K+,Mg2+,Ca2+,
strong acids,
thiols,
zwitterionic buffers,
nonionic buffers
Temperature RT RT
 

References:

Rohringer, R. and Holden, D.W. (1985) Protein blotting: detection of proteins with colloidal gold, and of glycoproteins and lectins with biotin-conjugated and enzyme probes. Anal Biochem 144, 118-127. ( or PubMed [PMID: 2984955] )

P.K. Smith et al. (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150, 76-85. ( or PubMed [PMID: 3843705] )

Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248-254. ( or PubMed [PMID: 942051] )

Schaffner W, Weissmann C. (1973) A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem 56(2), 502-14. ( or PubMed [PMID: 4128882] )

Lowry, O.H. et al. (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193, 265-275. ( or PubMed [PMID: 14907713] )

Lubran M.M. (1978) The measurement of total serum proteins by the Biuret method. Ann Clin Lab Sci 8(2), 106-10. ( or PubMed [PMID: 345944] )		  
 

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