This is the basis of biuret test widely used for identification of proteins and amino acids. If the purple color doesn’t appear, we are sorry to say that your test is negative. Aim: To estimate the protein using Biuret method. Since all proteins contain the peptide bond, the method is fairly specific and there is little interference from other compounds. Get a printable copy (PDF file) of the complete article (1.2M), or click on a page image below to browse page by page. Estimation of Protein by the Biuret Method Principle This is the most commonly used method based on the fact that the - CO - NH (peptide) group of proteins form a purple complex with copper ions in an alkaline medium. Some substances like urea and biuret interfere because they … Determination of Plasma Protein by Biuret Method Results: After performing the first two steps in the test, if the color of the solution turns to purple, it means that the proteins are present in the solution.

Reagents Required: 1. Biuret Reagent: Dissolve 3 g of copper sulphate (CuSO 4.5H 2O) and 9 g of sodium Upon complexation, a violet color is observed. Full text Full text is available as a scanned copy of the original print version. The Biuret Method, which is the most widely used method for total protein determination, relies on the complexation of Cu2+by the function groups involved with the peptide bond.
Aim: To estimate the protein using Biuret method.

Biuret is a compound formed by heating urea to 180° C. When biuret is treated with dilute copper sulfate in alkaline condition, a purple colored compound is formed. Links to PubMed are also available for Selected References. A minimum of two peptide bonds is needed for the complexation to occur.

The determination of protein concentration is an essential technique in all as pects of protein studies and proteomics. This test is given by compounds containing two or more peptide bond (CO-NH group). Principle: The –CO-NH- bond (peptide) in polypeptide chain reacts with copper sulphate in an alkaline medium to give a purple colour …

Principle: The –CO-NH- bond (peptide) in polypeptide chain reacts with copper sulphate in an alkaline medium to give a purple colour which can be measured at 540 nm. If possible, dilute unknowns to an estimated 1 to 10 mg/ml with buffer; a range of dilutions should be used if the actual concentration cannot be estimated. Prepare a reference tube with 1 ml buffer. Using 5 ml color reagent to 1 ml sample a recommended range is 0.5 to 20 mg protein. This lab activity is designed to teach students the principles behind a common protein estimation assay known as the Biuret Protein Assay.


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