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Bradford protein assay
About: Bradford protein assay is a research topic. Over the lifetime, 635 publications have been published within this topic receiving 239107 citations.
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TL;DR: It is demonstrated that as-prepared Au-Fe(3)O(4) nanoparticles can be modified with nitrilotriacetic acid molecules through Au-S interaction and used to separate proteins simply with the assistance of a magnet.
Abstract: In this article, we report the synthesis of bifunctional Au-Fe(3)O(4) nanoparticles that are formed by chemical bond linkage. Due to the introduction of Au nanoparticles, the resulting bifunctional Au-Fe(3)O(4) nanoparticles can be easily modified with other functional molecules to realize various nanobiotechnological separations and detections. Here, as an example, we demonstrate that as-prepared Au-Fe(3)O(4) nanoparticles can be modified with nitrilotriacetic acid molecules through Au-S interaction and used to separate proteins simply with the assistance of a magnet. Bradford protein assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis were performed to examine the validity of the separation procedure, and the phosphate determination method suggested that the as-separated protein maintained catalytic activity. This result shows the efficiency of such a material in protein separation and suggests that its use can be extended to magnetic separation of other biosubstances. Moreover, this synthetic strategy paves the way for facile preparation of diverse bifunctional and even multifunctional nanomaterials.
372 citations
TL;DR: Modified Scatchard and Hill analyses show that these high-affinity sites are few in number compared to the total number of dye-binding sites on a given protein.
306 citations
TL;DR: In this paper, the primary focus of this report is assay selection, emphasizing sample and buffer compatibility, and the fundamentals of generating protein assay standard curves and of data processing are considered, as are high-throughput adaptations of the more commonly used protein assays.
Abstract: Biochemical analysis of proteins relies on accurate quantification of protein concentration. Detailed in this appendix are some commonly used methods for protein analysis, e.g., Lowry, Bradford, bicinchoninic acid (BCA), UV spectroscopic, and 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) assays. The primary focus of this report is assay selection, emphasizing sample and buffer compatibility. The fundamentals of generating protein assay standard curves and of data processing are considered, as are high-throughput adaptations of the more commonly used protein assays. Also included is a rapid, inexpensive, and reliable BCA assay of total protein in SDS-PAGE sample buffer that is used for equal loading of SDS-PAGE gels. © 2016 by John Wiley & Sons, Inc.
291 citations
TL;DR: It is shown that under standard assay conditions, the ratio of the absorbance measurements at 590 nm and 450 nm is strictly linear with protein concentration, and a linear equation developed on the basis of mass action and Beer's law perfectly fits the experimental data.
Abstract: Determination of microgram quantities of protein in the Bradford Coomassie brilliant blue assay is accomplished by measurement of absorbance at 590 nm. This most common assay enables rapid and simple protein quantification in cell lysates, cellular fractions, or recombinant protein samples, for the purpose of normalization of biochemical measurements. However, an intrinsic nonlinearity compromises the sensitivity and accuracy of this method. It is shown that under standard assay conditions, the ratio of the absorbance measurements at 590 nm and 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 quantification down to 50 ng of bovine serum albumin. Furthermore, the interference commonly introduced by detergents that are used to create the cell lysates is greatly reduced by the new protocol. A linear equation developed on the basis of mass action and Beer's law perfectly fits the experimental data.
261 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