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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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Journal ArticleDOI
TL;DR: A novel easy-to-use homogeneous method utilizing two-photon excitation (TPX) for quantification of proteins or counting of eukaryotic cells in solution has been developed that tolerates interfering agents such as neutral detergents found in cell lysate samples even at high concentrations.
Abstract: A novel easy-to-use homogeneous method utilizing two-photon excitation (TPX) for quantification of proteins or counting of eukaryotic cells in solution has been developed. This highly sensitive technique is based on the adsorption competition between the sample and fluorescently labeled protein to micrometer-sized carboxylate modified polystyrene particles and detection of two-photon excited fluorescence. The adsorption of the labeled protein to the particles was detected as a distinct fluorescence on individual microparticles. Analyte protein or eukaryotic cells interacted with particle surface and reduced the adsorption of labeled protein to the particles resulting in a decrease of the fluorescence. The optimizations of assay conditions were performed separately for protein quantification and cell counting, and the principle of the method was confirmed with the fluorescence microscopy imaging. The protein quantification assay allowed the determination of picogram quantities (1.2 μg/L) of protein, and the cell counting assay allowed three cells in the sample with an average variation of approximately 10% in the signal. The protein assay sensitivity was more than 500-fold improved from the common most sensitive commercial methods. Moreover, the dynamic range of the assay was broad, approximately 4 orders of magnitude. The cell assay has sensitivity comparable to the most sensitive commercial method. The developed method tolerates interfering agents such as neutral detergents found in cell lysate samples even at high concentrations. The method is experimentally fairly simple and allows the expansion for the use of the TPX technology.

5 citations

Journal ArticleDOI
TL;DR: Urobilin has a strong confounding effect on the estimate of total protein concentrations obtained by BCA assay and thus this assay should not be used for urinary protein quantification, and it is recommended that the Bradford assay be used instead.

5 citations

Journal ArticleDOI
TL;DR: The CBQCA fluorescent assay for amines provides a simple assay for quantifying primary amine losses in dialysate fluid and values were substantially higher than previously reported amino acid losses.

5 citations

Journal ArticleDOI
TL;DR: In this article, a simple and effective method was devised to determine an enzyme incorporated into electropolymerized conductive polymer films, by combining a thin-layer EPC cell and the Bradford Coomassie colorimetric protein assay.
Abstract: A simple, rapid and effective way was devised to determine an enzyme incorporated into electropolymerized conductive polymer films, by combining a thin-layer electropolymerization cell and the Bradford Coomassie colorimetric protein assay. The method has been successfully applied to the determination of horseradish peroxidase (HRP) entrapped into pyrrole and 3-alkylsulfonate pyrrole copolymer films.

5 citations

Journal ArticleDOI
TL;DR: Two key aspects of assay accuracy were determining the dilutions of test sample that provided accurate quantitation (sample range), and performing spiking experiments at these dilutions to determine absence or presence of a "matrix" effect due to biological complexity of the sample.
Abstract: An Intracellular Adhesion Molecule I (ICAM-1) immunoassay from R and D Systems, and a Melanoma Inhibitory Activity (MIA) immunoassay from Roche Diagnostics were tested for accurate quantitation within complex biological substances such as cell lysates. Prior to assay, lysates of melanoma cells were treated with detergents to obtain soluble antigens. Maximum ICAM-1 and maximum MIA were detected after treatment using 0.8% Triton X-100. Two key aspects of assay accuracy were: 1) determining the dilutions of test sample that provided accurate quantitation (sample range), and 2) performing spiking experiments at these dilutions to determine absence or presence of a "matrix" effect due to biological complexity of the sample. A high degree of accuracy was found by diluting this particular cellular extract 50-fold prior to ICAM-1 assay, or only 5-fold prior to MIA assay. In addition, the bicinchoninic acid protein assay was analyzed to test the accuracy of protein quantitation of cellular lysates. Precision, limits of detection, and quantitation, robustness, linearity, and specificity also were tested for the immunoassays.

5 citations


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Performance
Metrics
No. of papers in the topic in previous years
YearPapers
20238
202212
202127
202021
201919
201822