Question

For each of the three assays (Bradford, Biuret, Folin-Ciocalteu) indicate what part of the protein reacts with the color forminh reagent and how does this specifically detect protein in a sample

Answer 1

Bradford method:

In Bradford reagents coomassie brilliant blue dye using identification of conc of total protein in Bradford in Bradford method in brilliant blue form of three types anionic , neutral, cationic in the form of acidic conditions of brilliant blue red convert to blue because proteins is present.

Binding protein is concerned of total protein we have to check Conc 595nm and plot the graph should be straight line.

Biuret method:

The biuret method is using for concentration of proteins because peptides bonds in presence of amino acids, biuret method used for detecting of peptides bonds in the presence of total solution. The biuret method copper with binds to nitrogen present in peptides of proteins.

The absorbance of Conc of biuret method 545nm we are performed.

Lowry's method (Folin-ciocalteu):

Lowry's method used for total protein concentration identification.oxidation of peptides bonds.

Lowry's method it's in biochemical analysis.

Lowry's method using of Folin-ciocalteu r(phospotungestic acid & phospho moylbidunic acid)

Lowry's method checking for absorbance 660 nm

Based on above 3 methods using for measuring of Conc of total protein.