During separation of amino acids using gel electrophoresis, can I use Biuret reagent or Ninhydrin.
Before answering i like to explain that the biuret test reagent is used to detect proteins in the the aqueous sample , a few drops of this reagent will turn the aqueous sample containing compounds with peptide bonds from pale to intense violet colour. . That is because proteins are made up of polypeptides, which in turn, are made of amino acids joined by peptide bonds. The longer the polypeptide chain is, the more peptide bonds there are, and therefore, the more intense the violet colour will be when biuret test is applied. It also follows that a pale violet or pinkish colour indicates shorter polypeptide chains or fewer peptide bonds. A negative result (lack of violet colour formation) may mean lack of protein, or the presence of free amino acids (without peptide bonds).
- according to above you cant use because :
1-the test need aqueous sample.
2-the color of reagent need presence polypeptide for acting.
Also the test, however, gives positive result to any compound containing two carbonyl groups attached to a nitrogen or carbon atom. Thus, it may not be completely protein-specific. Performing other tests may be necessary.
Thank you prof. Shadman, so can I use this reagent for detection of protein or polypeptide on Gel Electrophoresis?
I have used this test to detect peptides in eluates for preparitive electrophoresis experiments
- Badges
- Science method