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Author Topic: PROTEIN ASSAY METHOD: Protein estimation by taking Absorbance at 280nm  (Read 7474 times)
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Francis Umeoguaju
Expert in Bioscience Issues
Posts: 657

« on: April 10, 2010, 08:49:50 am »

This is a direct and fast method of protein analyses which involves taking the absorbance of a protein solution at 280nm.

Protein estimation by taking Absorbance at 280nm

This techniques is based on the absorption of aromatic amino acids (chiefly tryptophan and tyrosine) content of the protein at this wavelength. In most cases it is not used to quantitatively estimate protein concentration but it serves as quick method of comparing the protein of a sample with another sample especially in enzyme kinetics study.

For quantitative measure of protein concentration using the method, the absorbance of the sample is taken at both 260nm and 280nm to correct for the partial absorption of nucleic acid at 280nm. These is then substituted into calcap’s emperica formula
X = 1.45E280 – 0.74E260 where X is the concentration of protein in g/Litres. (Stroev et al 1989).

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