Showing papers on "Bradford protein assay published in 1985"
TL;DR: Bradford Coomassie brilliant blue G-250 protein-binding dye exists in three forms: cationic, neutral, and anionic; the binding behavior is attributed to Van der Waals forces and hydrophobic interactions.
616 citations
TL;DR: It was found that when sample buffer consisting of 9 M urea, 4 % Nonidet P‐40, 2 % Ampholines, and 2 % 2‐mercaptoethanol containing solubilized sample(s) was acidified prior to dilution, protein concentrations over a range of 0.5 to 50 μg could be reproducibly determined utilizing a modified Bradford assay.
Abstract: The concentration of protein actually solubilized in sample buffer in preparation for analysis by two-dimensional polyacrylamide gel electrophoresis cannot be directly determined by the Lowry, Biuret, or Bradford protein methods due to interference by the combinational effect of presence of urea, detergents, carrier ampholytes, and thiol compounds in sample solubilization buffers. Determinations of the actual amount of protein solubilized and applied to gels is required to accurately quantitatively and qualitatively evaluate second-dimension polypeptide maps. It was found that when sample buffer consisting of 9 M urea, 4 % Nonidet P-40, 2 % Ampholines, and 2 % 2-mercaptoethanol containing solubilized sample(s) was acidified prior to dilution, protein concentrations over a range of 0.5 to 50 μg could be reproducibly determined utilizing a modified Bradford assay. The modified assay generates two near-linear segments, one over the range < 0.5 to 5 μg total protein that permits the application of Beer's law and a second linear response encompassing 5 to 50 μg total protein. The assay did not tolerate presence of greater than 0.1 % sodium dodecyl sulfate but addition of sodium chloride and protamine sulfate did not adversely affect protein quantitation. The modified assay allows direct quantitation of protein solubilized in sample buffers containing urea, carrier ampholytes, nonionic detergents, and thiol compounds.
483 citations
TL;DR: The enhanced ability of this new method to tolerate high lipid levels without interference relative to several existing protein estimation methods is demonstrated.
225 citations
TL;DR: The dye-binding protein assay has been adapted for use with microtiter plates and a plate reader, and the response of nitrate reductase in the assay is about 70% of that of the standard protein, bovine serum albumin.
127 citations
TL;DR: A simple and rapid microassay for proteins utilizing the protein dye-staining procedure with a nitrocellulose (NC) filter is described, with good linearity between the optical density and the amount of protein obtained.
88 citations
TL;DR: Protein values determined by the dye-binding assay showed good agreement with those obtained by other procedures and several proteins immobilized on various matrices could be conveniently assayed.
18 citations
TL;DR: Large inconsistencies in replicating anaerobic runs for protein concentration appeared to be explained by noting that rising CO 2 bubbles may cause ‘foam fractionation’ of the proteins in the broth.
11 citations
TL;DR: A protein quantitation method which offers protein detection as low as 10 ng protein/ml and accurate quantitation as high as 30-100 ngprotein/ml, depending on the protein, has been designed.
6 citations
5 citations
TL;DR: A dye-impregnated test strip protein assay (Chemstrip-9) provides a rapid, visual assessment of membrane protein leakage and is sensitive to 200 mg/L of albumin in ultrafiltrates and do not react with as much as 4 mg/ L of phenytoin.
Abstract: Ultrafiltration techniques for the separation of bound and free ligand are dependent on minimal amounts of protein-bound drug leakage through the membrane devices. A dye-impregnated test strip protein assay (Chemstrip-9) provides a rapid, visual assessment of membrane protein leakage. The test strips are sensitive to 200 mg/L of albumin in ultrafiltrates and do not react with as much as 4 mg/L of phenytoin.
4 citations