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.

Assay of plant proteins with bicinchoninic acid for high resolution two-dimensional polyacrylamide gel electrophoresis.

Abstract: A method is described for estimating proteins in the same plant tissue sample that is solubilized for separation by two-dimensional polyacrylamide gel electrophoresis. The method uses a modified bicinchoninic acid (BCA) protein assay procedure and a modified standard urea solubilization buffer to estimate microgram values of unknown protein concentration, in the presence of 9 M urea and 4% Nonidet P-40, from a linear standard curve. A method for a quantitative determination of protein concentration by BCA in a sample containing 9 M urea and 4% Nonidet P-40 is also described. This method is effective for the determination of proteins in minute non-green and green plant tissue, and is especially designed for vegetative and floral shoot apices, and the primordia of inflorescences.

Degradation of Acid Red 27 by the Recombinant Flavin Reductase

Abstract: This paper focuses on the degradation of azo compound C.I. Acid Red 27 (AR27, amaranth) by the recombinant enzyme flavin reductase (FRE) from Citrobacter freundii strain A1. The enzyme was obtained via re-transformation of recombinant plasmid pET-43.1c(+)freBP containing the flavin reductase gene (fre) into E. coli NovaBlue. The plasmid was subsequently transformed into E. coli BL21(DE3)pLysS for overexpression of FRE fusion protein. Prior to that, the stability of fre gene was first verified using PCR amplification and also sequencing of the nucleotides and consequently compared with the original fre gene sequence from C. freundii A1. The protein was expressed as inclusion bodies and was isolated and refolded in order to obtain a properlyfolded and active flavin reductase. The activity and the protein yield were monitored using a modified flavin reductase assay and Bradford assay respectively. The active enzyme was further subjected to the degradation of AR27 and the degradation profile was constructed.

Determination of the Reaction Product of Glutaraldehyde and Amine Based on the Binding Ability of Coomassie Brilliant Blue

Abstract: Coomassie Brilliant Blue G-250 (CBBG) was bound to the reaction products of glutaraldehyde with various compounds containing amino group. The reaction products of acetaldehyde and malonaldehyde with the amines did not bind with CBBG, indicating that the binding was specific for the product of glutaraldehyde. The dye binding products rapidly formed in alkaline pH. Gel filtration chromatography indicated that a reaction product of more than 2 kDa has a dye- binding ability. The reaction product of glutaraldehyde with 6-aminohexanoic acid was separated with SP-Sephadex. CBBG was bound to the first eluted fraction from the column with highest affinity. The absorbance at 595nm by CBBG binding was linearly related to this lyophilized fraction at a concentration of up to 250μg/ml, suggesting its applicability to the determination of the reaction product of glutaraldehyde and amine. The present method should contribute to our understanding of the chemistry of glutaraldehyde associated with diverse fields, such as enzyme technology and biochemistry.