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By Eraldo Antonini, Luigi Rossi-Bernardi, Emilia Chiancone (Eds.)

The severely acclaimed laboratory normal, Methods in Enzymology, is among the so much hugely revered courses within the box of biochemistry. given that 1955, every one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. The sequence comprises a lot fabric nonetheless suitable this day - really a vital e-book for researchers in all fields of existence sciences

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Containing the ACD anticoagulant. 9% NaCI, and the top layer of white cells is removed. 0, and dialyzed for E. C. Abraham, A. Reese, M. Stallings, and T. H. J. Huisman, Hemoglobin 1, 27 (19761977). 97 A. L. Tan, A. DeYoung, and R. W. Noble, J. Biol. Chem. 247, 2493 (1972). R. G. Gillen and A. Riggs, J. Biol. Chem. 247, 6039 (1972). A. DeYoung, and R. W. Noble, private communication, 1980. l o o R. C. -Y. Tsay, Anal. Biochem. 54, 137 (1973). [2] PREPARATION OF MYOGLOBINS 29 1 hr against 1 liter of the same buffer.

20, 53 (1980). 4 R. L. Garlick, A m . Zool. 20, 69 (1980). R. E. Weber, A m . Zool. 20, 79 (1980). 6 j. Bonaventura and C. Bonaventura, in "Animals and Environmental Fitness" (R. ), p. 157. Pergamon, Oxford, 1980. METHODS IN ENZYMOLOGY, VOL. 76 Copyright © 1981 by Academic Press, Inc. All fights of reproduction in any form reserved. ISBN 0-12-181976-0 44 HEMOGLOBINS AND MYOGLOBINS [3] It has been shown 12 that one of the polypeptide chains of the erythrocruorin from the earthworm Lumbricus is homologous to the chains of other members of the globin family.

Reduction is complete within 20 min, and solutions of oxyhemoproteins so prepared are stable for about 1 week. Stock enzyme solutions or suspensions are used as purchased without further dilution. 38 Myoglobin may also be reduced with dithionite. 0. A layer about 1 cm high, of an anaerobically prepared solution of dithionite, 10-100 mM, in the same buffer is carefully pipetted above the surface of the Sephadex bed, beneath a protecting column of anaerobic buffer, and is run into the column. A solution of ferric myoglobin is next applied to the column.

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