Elsevier

Clinica Chimica Acta

Volume 33, Issue 2, July 1971, Pages 379-386
Clinica Chimica Acta

A NADH-dependent transketolase assay in erythrocyte hemolysates

https://doi.org/10.1016/0009-8981(71)90496-7Get rights and content

Abstract

A simple and specific method for the assay of transketolase activity in erythrocyte hemolysates based on the measurement of NADH consumption is described. The test conditions are discussed and the results are compared with a colorimetric method. Normal values are given for the transketolase activity and the thiamine pyrophosphate effect.

References (25)

  • M. Brin

    J. Nutr.

    (1962)
  • M. Brin et al.

    J. Biol. Ctiem.

    (1958)
  • M. Brin et al.

    J. 'Nutr.

    (1960)
  • Z.Z. Ziporin et al.

    J. Nutr.

    (1965)
    Z.Z. Ziporin et al.

    J. Nutr.

    (1965)
  • M.S. Bamji

    Amer. J. Clin. Nutr.

    (1970)
  • M. Brin et al.
  • T. Markkanen et al.

    Amer. J. Med. Sci.

    (1966)
  • B.L. Horecker et al.

    J. Biol. Chem.

    (1953)
  • E.J. Van Kampen et al.

    Clin. Chim. Acta

    (1961)
  • R. Szok et al.

    Advan. Protein Chem.

    (1960)
  • A.H. Brown

    Arch. Biochem.

    (1946)
  • M. Brin et al.
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