Determination of levels of glycolytic intermediates and nucleotides in platelets by pulse-labeling with [32P]orthophosphate
✍ Scribed by Holm Holmsen; Carol A. Dangelmaier; Jan-Willem N. Akkerman
- Book ID
- 102986895
- Publisher
- Elsevier Science
- Year
- 1983
- Tongue
- English
- Weight
- 996 KB
- Volume
- 131
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A method is presented for the separation and quantificrttion of 32P-labeled carbohydrates and nucleotides in blood platelets which have been pulse-labeled with [32P]orthophosphate. The procedure is based on twodimensional paper chromatography, identification of the spots by radioautography and enzymatic methods, and quantitation of 32P radioactivity by liquid scintillation counting. The data show that 32P is homogeneously distributed among the compounds studied so that the total radioactivity is proportional to the levels of these compounds in the metabolic compartment of the cells. Thus, this method provides a sensitive and accurate means to evaluate phosphorylated intermediates in glycolysis and nucleotide metabolism and to assess the transfer of energy-rich phosphate groups between these pathways in particular.