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Preparation of Tissues for Analysis

  • Janet V. Passonneau
  • Oliver H. Lowry
Protocol
Part of the Biological Methods book series (BM)

Abstract

Often the preparative steps in a tissue analysis are the most critical. In the case of a metabolite assay, the most hazardous period is usually between the moment the blood supply is cut off and the moment enzyme action is finally stopped. In the case of an enzyme, with a few important exceptions, there is not likely to be much change for many minutes or even hours. Instead, the biggest problem may be to render the enzyme fully accessible for assay, or to prevent loss during or after homogenization. In a few cases, if the objective is to determine the state of activity as it was in vivo, the problem may be to prevent a specific activity change of the enzyme (e.g., the conversion of phosphorylase b to a, and so on).

Keywords

Freeze Tissue Pyridine Nucleotide Tissue Fluorescence Enzymatic Cycling Metabolite Assay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Richter, D. and Dawson, R. M. C. (1948) Brain metabolism in emotional excitement and in sleep. Amer. J. Physiol. 154, 73–79.PubMedGoogle Scholar
  2. 2.
    Wollenberger, A., Ristau, O., and Schoffa, G. (1960) Eine einfache technik der extrem schnellen abkühling grösserer gewebestücke. Pflueger’s Arch. Gesamte Physiol. Menschen Tiere. 270, 399–412.CrossRefGoogle Scholar
  3. 3.
    Kerr, S. E. and Ghantus, M. (1937) The carbohydrate metabolism of the brain. III. On the origin of lactic acid. J. Biol. Chem. 117, 217–225.Google Scholar
  4. 4.
    Ferrendelli, J. A., Gay, M. H., Sedgwick, W. G., and Chang, M. M. (1972) Quick-freezing of the murine CNS: Comparison of regional cooling rates and metabolite levels when using liquid nitrogen or Freon-12. J. Neurochem. 19, 979–987.PubMedCrossRefGoogle Scholar
  5. 5.
    Nelson, S. R., Lowry, O. H., and Passonneau, J. V. (1966) in Head Injury Conference Proceedings (Caveness, W. F. and Walker, A. E., eds.), Lippincott, Philadelphia, PA, p. 444.Google Scholar
  6. 6.
    Bessey, O. H., Lowry, O. H., and Love, R. H. (1949) The fluorometric measurement of the nucleotides of riboflavin and their concentration in tissues. J. Biol. Chem. 180, 755–769.PubMedGoogle Scholar
  7. 7.
    Burch, H. B., Bradley, M. E., and Lowry, O. H. (1967) The measurement of triphosphopyridine nucleotide and the role of hemoglobin in producing erroneous triphosphopyridine nucleotide values. J. Biol. Chem. 242, 4546–4554.PubMedGoogle Scholar

Copyright information

© The Humana Press Inc. 1993

Authors and Affiliations

  • Janet V. Passonneau
    • 1
  • Oliver H. Lowry
    • 2
  1. 1.WashingtonUSA
  2. 2.Washington UniversitySt. LouisUSA

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