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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).
By using the Bradford protein assay, one can avoid all of these complications by simply mixing the protein samples with the Coomassie brilliant blue G-250 dye (Bradford reagent) and measuring their absorbances at 595 nm, which is in the visible range [8] and may be accurately measured by the use of a mobile smartphone camera. [9]
Protein methods are the techniques used to study proteins. There are experimental methods for studying proteins (e.g., for detecting proteins, for isolating and purifying proteins, and for characterizing the structure and function of proteins, [ 1 ] often requiring that the protein first be purified).
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 alternative method for label free protein quantification in clear liquid is cuvette-based SPR technique, that simultaneously measures the refractive index ranging 1.0 to 1.6 nD and concentration of the protein ranging from 0.5 μL to 2 mL in volume. This system consists of the calibrated optical filter with very high angular resolution and ...
This method is able to detect as low as 25 μg/ml and up to 2000 μg/ml of protein in a 65 ul sample, using standard protocol. This method may be preferred for samples containing detergents or other reducing agents. This method has a fast detection speed and low protein-to-protein variability in comparison to the BCA or Coomassie (Bradford ...
The Kjeldahl method's universality, precision and reproducibility have made it the internationally recognized method for estimating the protein content in foods and it is the standard method against which all other methods are judged. It is also used to assay soils, waste waters, fertilizers and other materials.
The Warburg–Christian method is an ultraviolet spectroscopic protein and nucleic acid assay method based on the absorbance of UV light at 260 nm and 280 nm wavelengths. Proteins generally absorb light at 280 nanometers due to the presence of tryptophan and tyrosine .