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J Biolumin Chemilumin. 1991 Apr-Jun;6(2):123-9.

A luminometric method for the determination of ATP and phosphocreatine in single human skeletal muscle fibres.

Author information

1
Department of Clinical Chemistry II, Huddinge University Hospital, Sweden.

Abstract

A sensitive method for the analysis of ATP and phosphocreatine (PCr) in single human skeletal muscle fibres is described. Muscle tissue was freeze-dried and single fibres were dissected free with the aid of low-power microscopy. The fibres were then extracted in trichloroacetic acid and neutralized with KHCO3. The assay is based on the continuous monitoring of light produced as a result of ATP degradation in the firefly luciferase reaction. PCr is measured as the amount of ATP formed in the creatine kinase reaction. The coefficient of variation was less than 4% for both ATP and PCr determination. The amount of tissue required for the assay is approximately 0.5 microgram (dry weight). The assay showed good agreement with spectrophotometric and high-performance liquid chromatographic (HPLC) measurements made upon extracts of whole muscle tissue.

PMID:
1882705
DOI:
10.1002/bio.1170060210
[Indexed for MEDLINE]

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