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Bradford protein assay

About: Bradford protein assay is a(n) research topic. Over the lifetime, 635 publication(s) have been published within this topic receiving 239107 citation(s).


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Journal ArticleDOI
TL;DR: This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr with little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose.
Abstract: A protein determination method which involves the binding of Coomassie Brilliant Blue G-250 to protein is described. The binding of the dye to protein causes a shift in the absorption maximum of the dye from 465 to 595 nm, and it is the increase in absorption at 595 nm which is monitored. This assay is very reproducible and rapid with the dye binding process virtually complete in approximately 2 min with good color stability for 1 hr. There is little or no interference from cations such as sodium or potassium nor from carbohydrates such as sucrose. A small amount of color is developed in the presence of strongly alkaline buffering agents, but the assay may be run accurately by the use of proper buffer controls. The only components found to give excessive interfering color in the assay are relatively large amounts of detergents such as sodium dodecyl sulfate, Triton X-100, and commercial glassware detergents. Interference by small amounts of detergent may be eliminated by the use of proper controls.

214,383 citations

Journal ArticleDOI
TL;DR: Under the new conditions there is direct proportionality between absorbance at 650 nm and weight of protein within the range 15–110 μg.
Abstract: 1. 1. The Lowry, Rosebrough, Farr, and Randall method for protein assay has been modified so as to give a higher color yield with bovine serum albumin and with five other pure proteins. 2. 2. Under the new conditions there is direct proportionality between absorbance at 650 nm and weight of protein within the range 15–110 μg.

5,067 citations

Book ChapterDOI
TL;DR: A rapid and accurate method for the estimation of protein concentration is essential in many fields of protein study, but is susceptible to interference from a wide range of compounds commonly present in biological extracts.
Abstract: A rapid and accurate method for the estimation of protein concentration is essential in many fields of protein study. The Lowry method ( Chapter 1 in vol. 1 of this series) has been widely used, but is susceptible to interference from a wide range of compounds commonly present in biological extracts. Although interference can be avoided by trichloracetic acid precipitation of the protein prior to assay, this lengthens the procedure.

1,318 citations

Journal ArticleDOI
TL;DR: Different sensitivity was observed for each assay with the neutral red and the MTT assay being the most sensitive in detecting cytotoxic events compared to the LDH leakage and the protein assay.
Abstract: The aim of this study was to compare four in vitro cytotoxicity assays and determine their ability to detect early cytotoxic events. Two hepatoma cell lines, namely HTC and HepG2 cells, were exposed to cadmium chloride (0-300 microM) for 3, 5 and 8 h. Following exposure to the toxic metal cytotoxicity was determined with the lactate dehydrogenase leakage assay (LDH), a protein assay, the neutral red assay and the methyl tetrazolium (MTT) assay. In HTC cells no toxicity was observed for any incubation period when the LDH leakage, the MTT and the protein assay were employed whereas the neutral red assay revealed early cytotoxicity starting after incubation of HTC cells with CdCl(2) for 3 h. In the case of HepG2 cells the MTT assay reveals cytotoxicity due to CdCl(2) exposure after 3 h whereas no such effect is seen with the other three assays. Following 5 h exposure of HepG2 cells to CdCl(2), toxicity is observed with the MTT assay at lower concentrations compared to the ones required for detection of toxicity with the LDH leakage and the neutral red assay. In conclusion different sensitivity was observed for each assay with the neutral red and the MTT assay being the most sensitive in detecting cytotoxic events compared to the LDH leakage and the protein assay.

1,177 citations

Journal ArticleDOI
TL;DR: Under standard assay conditions, the ratio of the absorbances, 590 nm over 450 nm, is strictly linear with protein concentration, and a linear equation that perfectly fits the experimental data is provided on the basis of mass action and Beer's law.
Abstract: Determination of microgram quantities of protein in the Bradford Coomassie brilliant blue assay is accomplished by measurement of absorbance at 590 nm. However, an intrinsic nonlinearity compromises the sensitivity and accuracy of this method. It is shown that under standard assay conditions, the ratio of the absorbances, 590 nm over 450 nm, is strictly linear with protein concentration. This simple procedure increases the accuracy and improves the sensitivity of the assay about 10-fold, permitting quantitation down to 50 ng of bovine serum albumin. Furthermore, protein assay in presence of up to 35-fold weight excess of sodium dodecyl sulfate (detergent) over bovine serum albumin (protein) can be performed. A linear equation that perfectly fits the experimental data is provided on the basis of mass action and Beer's law.

938 citations

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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
202127
202021
201919
201822
201710
201627