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Coomassie blue protein dye-binding assays measure formation of an insoluble protein-dye complex

โœ Scribed by Thomas Marshall; Katherine M. Williams

Publisher
Elsevier Science
Year
1992
Tongue
English
Weight
216 KB
Volume
204
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

The solubility of the protein-Coomassie brilliant blue (CBB) complex formed upon Bradford (Anal. Biochem. 72, 248-254, 1976) or Sedmak and Grossberg (Anal. Biochem. 79, 544-552, 1977) protein assay has been investigated by centrifugation or filtration of the assay mix within 10 min of adding dye reagent. The results show complete loss of color yield in the respective supernates and filtrates. This indicates that the protein-CBB complexes are insoluble at the time of absorbance measurement. Protein solubility in the dye reagent may dictate the relative response of the assay to an individual protein and the requirement for macromolecular structure.

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The direct measurement of protein in cell suspensions using the Coomassie brilliant blue dyebinding assay demonstrated markedly lower values compared to those obtained with the Lowry assay. It is shown that the addition of small amounts of Triton X-100 or NaOH to the cell suspensions prior to additi