Adaptation of the Bradford protein assay to membrane-bound proteins by solubilizing in glucopyranoside detergents

摘要: A procedure was developed for the quantitation of solubilized proteins using Bradford assay in presence glucopyranoside detergents. These detergents membrane-bound with minimal background absorbance at 595 nm. Absorbance 650 nm also low, indicating that these do not significantly stabilize neutral species Coomassie brilliant blue G-250 produces interference other Hexyl-beta-D-glucopyranoside produced less than did larger glucopyranosides, and increase its dye reagent related linearly to concentration from 0 2%. by protein increased up 0.2% hexyl-beta-D-glucopyranoside (final reagent) then remained stable 1%, concentrations this detergent allowed react completely reagent. Standard curves several were similar absence or 0.1-0.5% hexyl-beta-D-glucopyranoside. The both soluble hexyl-, heptyl-, octyl-beta-D-glucopyranoside. Estimates agreed estimates obtained Lowry quantitative amino acid analysis. This requires no extra steps; thus, it is as rapid convenient original assay.