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Protein concentrations in cell-free extracts can be determined quickly and accurately using the measured absorbances at 230 and 260 nm. The choice of these two wavelengths, rather than those (280 and 260 nm) used by
Warburg and Christian [(1942)
Biochem. Z.
310, 384–421]
, decreases interference by nucleic acids and results in sensitivity equivalent to that of the method of
Lowry, Rosebrough, Farr, and Randall [(1951)
J. Biol. Chem.
193, 265–275]
. The
230
260
method is less dependent on changes in the amino acid composition of the protein(s) being measured than is the
280
260
method. Using buffered solutions of crystalline bovine serum albumin and yeast RNA as standards, we derived the following equation: Protein concentration (
μg/ml) = 183
A
230 − 75.8
A
260, where
A
230 and
A
260 are the absorbances at 230 and 260 nm.