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The method combines the reactions of copper ions with the peptide bonds under alkaline conditions (the Biuret test) with the oxidation of aromatic protein residues. The Lowry method is based on the reaction of Cu +, produced by the oxidation of peptide bonds, with Folin–Ciocalteu reagent (a mixture of phosphotungstic acid and phosphomolybdic acid in the Folin–Ciocalteu reaction).
The Bradford protein assay (also known as the Coomassie protein assay) was developed by Marion M. Bradford in 1976. [1] It is a quick and accurate [2] spectroscopic analytical procedure used to measure the concentration of protein in a solution. The reaction is dependent on the amino acid composition of the measured proteins.
Bicinchoninic acid assay (BCA assay): Detection down to 0.5 μg/mL; Lowry Protein assay: Detection in the range of 0.01–1.0 mg/mL; Fluorescamine: Quantifies proteins and peptides in solution if primary amine are present in the amino acids; Amido black: Detection in the range of 1-12 μg/mL; Colloidal gold: Detection in the range of 20 - 640 ng/mL
BCA protein assay in a 96 well plate. The bicinchoninic acid assay (BCA assay), also known as the Smith assay, after its inventor, Paul K. Smith at the Pierce Chemical Company, [1] now part of Thermo Fisher Scientific, is a biochemical assay for determining the total concentration of protein in a solution (0.5 μg/mL to 1.5 mg/mL), similar to Lowry protein assay, Bradford protein assay or ...
An assay (analysis) is never an isolated process, as it must be accompanied with pre- and post-analytic procedures. Both the communication order (the request to perform an assay plus related information) and the handling of the specimen itself (the collecting, documenting, transporting, and processing done before beginning the assay) are pre-analytic steps.
As of September 2015, his 1951 paper in the Journal of Biological Chemistry [5] describing the protein assay was still the most-highly cited paper in history, with more than 310,000 citations, [6] although Lowry stated it was not the most important paper he had ever written.
Bradford was born October 28, 1946, in Rome, Georgia, US, and received his B.A. from Shorter College there in 1967. [1] In 1971 he married Janet Holliday. [1] [8] He obtained his Ph.D. in biochemistry from the University of Georgia in 1975, and his use of the Coomassie Brilliant Blue G-250 dye to detect proteins, which became known as the Bradford assay, was patented in 1976.
A calibration curve plot showing limit of detection (LOD), limit of quantification (LOQ), dynamic range, and limit of linearity (LOL).. In analytical chemistry, a calibration curve, also known as a standard curve, is a general method for determining the concentration of a substance in an unknown sample by comparing the unknown to a set of standard samples of known concentration. [1]