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Pedersen, "The Ultracentrifuge," p. 358. Oxford Univ. Press (Clarendon), London and New York, 1940. s j. Roche, in "Studies in Comparative Biochemistry" (K. A. ), p. 62. Pergamon, Oxford, 1965. 4 R. E. Weber, Comp. Biochem. Physiol. 35, 179 (1970). 5 L. Waxman, J. Biol. Chem. 246, 7318 (1971). 6 j. B. Swaney and I. M. Klotz, Arch. Biochem.

J. Bergersen, C. A. Appleby, and G. L. Turner, J. Biol. Chem. 249, 4057 (1974). 73 Q. H. Gibson and M. H. Smith, Proc. R. Soc. London, Ser. B 163, 206 (1965). 4, containing 1 mM EDTA, and the supernatant solution was separated by centrifugation. 2 and fractionated by the addition of solid ammonium sulfate. The fraction precipitating between 56 and 80% saturation was retained. 0, and was separated from high molecular weight substances by chromatography on a column of Sephadex G-75 (5 × 52 cm) in the same buffer.

J. 133, 725 (1973). s4 D. Keilin, Nature (London) 190, 717 (1961). so J. S. Haldane, J. Physiol. (London) 25, 230 (1900). so S. Tomita, Y. Enoki, M. Santa, H. Yoshida, and Y. Y a s u m i t s u , J . Nara Med. Assoc. 19, 1 (1968). 24 HEMOGLOBINS AND MYOGLOBINS [1] moglobin content of a dilute oxy- or carbonylhemoglobin solution. Their method depends on conversion to the CNMetHb form and is independent of pH. A few crystals of KCN are added to 5 ml of the hemoglobin solution. Let stand 20 min at 20-25 ° (not at 38° as originally describedsn), then measure the absorbance, A1, at one of the following wavelengths: if HbO~, at 542 nm or 576 nm; if HbCO, at 540 or 570 nm.

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