Elsevier

Analytical Biochemistry

Volume 82, Issue 2, October 1977, Pages 362-371
Analytical Biochemistry

A new spectrophotometric assay for protein in cell extracts

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Abstract

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 230260 method is less dependent on changes in the amino acid composition of the protein(s) being measured than is the 280260 method. Using buffered solutions of crystalline bovine serum albumin and yeast RNA as standards, we derived the following equation: Protein concentration (μg/ml) = 183 A230 − 75.8 A260, where A230 and A260 are the absorbances at 230 and 260 nm.

References (29)

  • W.E. Groves et al.

    Anal. Biochem

    (1968)
  • R.K. Scopes

    Anal. Biochem

    (1974)
  • O.H. Lowry et al.

    J. Biol. Chem

    (1951)
  • B. Ehresmann et al.

    Anal. Biochem

    (1973)
  • I.M. Kolthoff et al.

    Anal. Biochem

    (1965)
  • R.E. Canfield

    J. Biol. Chem

    (1963)
  • A.G. Gornall et al.

    J. Biol. Chem

    (1949)
  • T. Lindahl et al.
  • R.W. Holley
  • K.S. Kirby
  • G. Felsenfeld et al.

    J. Mol. Biol

    (1965)
  • H.H. Ohlenbusch et al.

    J. Mol. Biol

    (1967)
  • K. Miura

    Biochim. Biophys. Acta

    (1962)
  • V. Massey et al.

    J. Biol. Chem

    (1965)
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    A preliminary report has been presented before the 59th annual meeting of the American Society for Experimental Biology (1975) Fed. Proc.34, 546.

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