Refinement of the Coomassie brilliant blue G assay for quantitative protein determination.
Minor modifications of the Bradford method [1976) Anal. Biochem. 72, 248-254) increase the sensitivity of the Coomassie brilliant blue G microassay for protein quantitation. This is reached by selecting the proper dye source, by alterations in the dye, ethanol, and phosphoric acid concentrations, and by including Triton X-100 in the reaction mixture. The modified assay conditions allow proteins to be detected above 0.2 micrograms and from a solution above 0.4 micrograms/ml. The extinction coefficient of the dye-protein complex was 0.144 +/- 0.010 for 1 microgram bovine serum albumin in the final reaction mixture of 1 ml. In general, the modified procedure exhibits the same specificity, advantages, and drawbacks as described for the original Bradford assay.
Löffler
B M
BM
Max-Planck-Institut für Experimentelle Medizin, Göttingen, Federal Republic of Germany.
Kunze
H
H
D017830:Octoxynol; D010756:Phosphoric Acids; D011092:Polyethylene Glycols; D011506:Proteins / Q000032:analysis*; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine / Q000032:analysis; D013053:Spectrophotometry
D010756:Phosphoric Acids; D011506:Proteins; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols; D017830:Octoxynol; C030242:phosphoric acid; C004692:coomassie Brilliant Blue