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Encyclopedia > Biuret reagent

The Biuret reagent is made of potassium hydroxide (KOH) and copper (II) sulfate (CuSO4), together with potassium sodium tartrate (KNaC4H4O6·4H2O). The blue reagent turns violet in the presence of proteins, and changes to pink when combined with short-chain polypeptides. The potassium hydroxide does not participate in the reaction at all, but is merely there to provide an alkaline medium so that the reaction can take place. The chemical compound potassium hydroxide, (KOH) sometimes known as caustic potash, potassa, potash lye, and potassium hydrate, is a metallic base. ... Copper (II) sulfate (CuSO4) is the most common copper salt, made by the action of sulfuric acid on the base copper oxide. ... Potassium sodium tartrate is a double salt first prepared (in about 1675) by an apothecary, Pierre Seignette, of La Rochelle, France. ... A representation of the 3D structure of myoglobin, showing coloured alpha helices. ... Peptides are the family of molecules formed from the linking, in a defined order, of various amino acids. ... The common (Arrhenius) definition of a base is a chemical compound that either donates hydroxide ions or absorbs hydrogen ions when dissolved in water. ...


The reagent is commonly used in a biuret protein assay, a colorimetric assay used to determine protein concentration--such as UV-VIS at wavelength 560nm (to detect the Cu2+ ion). A representation of the 3D structure of myoglobin, showing coloured alpha helices. ... An assay is a procedure where the concentration of a component part of a mixture is determined. ... A colorimeter is device used to measure the absorbance of a specific solution. ... For other uses, see Concentration (disambiguation). ... Ultraviolet-Visible Spectroscopy or Ultraviolet-Visible Spectrophotometry (UV/ VIS) involves the spectroscopy of photons (spectrophotometry). ...


The Biuret Test

Main Article: Biuret test

The principle underlying the test can be demonstrated with the chemical compound biuret which, just as proteins, is able to complex copper (II) ions. The biuret assay does not in fact use biuret, but is so named as it detects the peptide bond between the urea molecules or between amino acids. Biuret test is used for determining the amount of protein in a food sample. ... Biuret is a chemical compound with the chemical formula C2O2N3H5. ... Synthesis of copper(II)-tetraphenylporphine, a metal complex, from tetraphenylporphine and copper(II) acetate monohydrate. ... For other uses, see Copper (disambiguation). ... This article is about the electrically charged particle. ... Biuret is a chemical compound with the chemical formula C2O2N3H5. ... A peptide bond is a chemical bond that is formed between two molecules when the carboxyl group of one molecule reacts with the amino group of the other molecule, releasing a molecule of water (H2O). ...


Not all biuret tests actually require the Biuret reagent. Rather, the term "biuret test" is a generic term for the testing of proteins by using copper (II) sulfate solution in an alkaline environment. Thus, the process of testing for proteins in a solution by first adding a small amount of sodium hydroxide, and then adding copper (II) sulfate drop by drop, is also known as a biuret test. Flash point Non-flammable. ...


References

  • Chemical Reagents

  Results from FactBites:
 
Protein determination by the biuret method (445 words)
The principle of the biuret assay is similar to that of the Lowry, however it involves a single incubation of 20 min.
The biuret is a good general protein assay for batches of material for which yield is not a problem.
The reported sensitive range, using 9 ml reagent to 1 ml sample, is from 1 to 10 mg protein.
Spectrophotometric Protein Assays (328 words)
The latter three are for the biuret assay and the former are for the rest.
Biuret: Mix 0.50 mL of protein with 2.50 mL of biuret reagent and measure the absorbance at 540 nm.
Bradford: Mix 0.25 mL of protein with 2.5 mL of Bradford reagent and measure the absorbance at 595 nm.
  More results at FactBites »


 
 

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