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Determination of the Extinction Coefficient for a Protein
Use the following formula to determine the extinction coefficient at 205 nm:
E(205 nm) = 27 + 120 x (A280 divided by A205)
The reading at 205 nm must be multiplied by the dilution factor before using the formula.
Next, determine protein concentration:
Protein concentration (M) = A205 divided by E(205 nm)
You can now determine the extinction coefficient for 280 nm:
E(280 nm) = concentration (M) divided by A280
Comments
An abnormal phenylalanine content will throw off the result considerably. The accuracy
of the technique depends on an average amino acid composition.
References
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Scopes, RK. Analytical Biochemistry 59: 277. 1974.
Stoscheck, CM. Quantitation of Protein. Methods in Enzymology 182: 50-69.
1990.
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