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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 Article
TL;DR: It was demonstrated that elution of the protein was very rapid, and changes of the extraction buffer volume had no appreciable effect on the extraction efficiency, and comparison experiments of the four assay methods revealed that Ninhydrin method was significantly interfered by low molecular weight substances other than protein and that Bradford method suffered from the poor reproducibility.
Abstract: Recently type I hypersensitivity reactions caused by latex gloves have been reported. The protein from latex gloves is considered to be responsible for this allergy. We investigated the extraction conditions and the assay methods for the purpose of the colorimetric determination of the total protein eluted from latex gloves. In a series of experiments, it was demonstrated that elution of the protein was very rapid, and changes of the extraction buffer volume (1.7-20 ml/g) had no appreciable effect on the extraction efficiency. Moreover, comparison experiments of the four assay methods revealed that Ninhydrin method was significantly interfered by low molecular weight substances other than protein and that Bradford method suffered from the poor reproducibility. On the other hand, Lowry method and Bicinchoninic acid (BCA) method gave relatively good results. Based on these experimental results, we established a typical procedure for quantitative analysis of the total protein as follows. Two grams of latex gloves specimens are extracted with 10 ml portions of phosphate buffered saline pH 7.4 (PBS) at room temperature for two hours, and then the resulting solution is assayed by BCA method. According to this procedure, commercially available ten kinds of latex gloves were analyzed. We found that the amount of the extractable protein was considerably varied with the products.

3 citations

Journal ArticleDOI
TL;DR: The results showed that the presence of surfactant was very useful in preventing the initial adsorption of EPO on the walls of vials and in minimizing protein aggregation.
Abstract: The aim of this study was to optimize the formulation of erythropoietin (EPO) using amino acids instead of human serum albumin (HSA) and to evaluate its in vivo stability in order to avoid the risk of viral contamination and antigenicity. Different EPO formulations were developed in such a way as to allow studying the effects of amino acids and surfactants on the EPO stability profile. The main techniques applied for EPO analysis were ELISA, Bradford method, and SDS gel electrophoresis. The in vivo stability was evaluated in a Balb-c mouse animal model. The results showed that the presence of surfactant was very useful in preventing the initial adsorption of EPO on the walls of vials and in minimizing protein aggregation. Amino acid combinations, glycine with glutamic acid, provided maximum stability. Formulation F4 (containing glycine, glutamic acid and Tween 20) showed minimum aggregation and degradation and in vivo activity equivalent to commercially available HSA-stabilized EPO (Eprex®).

3 citations

Journal ArticleDOI
TL;DR: The results showed that serum protein adsorption and complement activation were augmented for nanoparticles with a larger size below 400 nm, and nanoparticles in the nanometer and submicrometer range were phagocytosed more readily than either smaller or larger particles.
Abstract: The purpose of this study is to evaluate the effect of particle size on serum protein opsonization and in vitro macrophage uptake of polyethyleneglycol modified poly (D, L-lactide-co-glycolide) nanoparticles (PEG-PLGA-NPs). PEG-PLGA-NPs were prepared by modified-spontaneous emulsification solvent diffusion (modified-SESD) method. Serum protein adsorptions to PEG-PLGA-NPs were evaluated by bicinchoninic acid (BCA) protein assay and enzyme-linked immunosorbent assay (ELISA). Complement activation was also investigated by ELISA for complement fragments iC3b. Uptake of PEG-PLGA-NPs by macrophages was measured by fluorescence spectrometer. The results showed that serum protein adsorption and complement activation were augmented for nanoparticles with a larger size below 400 nm. Phagocytosis of PEG-PLGA-NPs by murine peritoneal macrophages involved serum-independent and serum-dependent phagocytosis. Serum-independent phagocytosis decreased, while serum-dependent phagocytosis increased with the increase of particle size in the nanometer and submicrometer range. Consequently, nanoparticles with size of about 400 nm were phagocytosed more readily than either smaller or larger particles

3 citations

Patent
05 Oct 2016
TL;DR: In this paper, a whole-course C-reactive protein assay kit is described, which includes an M reagent, an R1 reagent and an R2 reagent.
Abstract: The invention discloses a whole-course C-reactive protein assay kit and an application thereof. The kit includes the following components: an M reagent, an R1 reagent, an R2 reagent, an alkaline pre-treatment solution, a pre-excitation agent and an excitation agent. The M reagent includes 0.5-1 mg/ml of magnetic particles coated by a first antibody, 0.5-1% of casein and 0.5-1% of bovine serum albumin, wherein the solvent is a phosphate buffer solution being 7.0-8.0 in pH. The coating quantity of the first antibody is 5-15 [mu]g per mg of the magnetic particles. The R1 reagent is an HCl solution being 10-20 mM in concentration. The R2 reagent includes acridinium ester which is coated by a second antibody, 0.5-1% of casein and 0.5-1% of bovine serum albumin, wherein the solvent is a phosphate buffer solution being 7.0-8.0 in pH. The coating quantity of the second antibody is 5-15 [mu]g per ml of the acridinium ester. The alkaline pre-treatment solution is a NaOH solution being 20-50 mM in concentration. Strong alkali is added to the kit during reaction to damage the proteins, and then strong acid is added to neutralize the reaction system. The kit is 0.02-100 mg/l in detection range, and the reagent can reach the requirement of detection of the whole course of C-reactive protein.

3 citations


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