Miniature gel filtration: a simple and inexpensive method for the measurement of protein- and non-protein-bound cortisol: application to guinea pig plasma |
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| Affiliation: | 1. Department of Microbiology, Institute of Biomedical Science, University of São Paulo, Av. Prof Lineu Prestes, 1374, CEP 05508-000 São Paulo, SP, Brazil;2. Department of Chemistry. The University of Warwick, Coventry CV4 7AL, UK;1. Department of Physics, Chemistry and Biology, Linköping University, Sweden;2. European Molecular Biology Laboratory (EMBL), 71 Avenue des Martyres, 38000 Grenoble, France;3. ALPX S.A.S. 71 Avenue des Martyrs, 38000 Grenoble, France;4. Department of Biochemistry, Netherlands Cancer Institute, Amsterdam, the Netherlands;5. Oncode Institute, the Netherlands;6. Wallenberg Center for Molecular Medicine, Linköping University, Sweden |
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| Abstract: | - 1.1. A method is described for the accurate and rapid measurement of protein- and non-protein-bound cortisol by miniature gel filtration in small volumes of plasma, e.g. of rodents.
- 2.2. Binding of cortisol by guinea pig plasma proteins is strongly reduced at elevated temperature (4°C: 102 ± 12ng/ml; 40°C: 5 ± 2 ng/ml).
- 3.3. Incubation of guinea pig plasma with 1–5000 ng cortisol resulted in a dose-dependent increase in cortisol bound to proteins (specific binding by corticosteroid binding globulin: 230 ± 12 ng/ml).
- 4.4. Administration of 20 IU (1–24)ACTH induced a significant increase of total protein-bound and non-protein-bound cortisol.
- 5.5. Values reported in this study agree well with those of previous investigations, in which bound and non-bound glucocorticosteroids were separated by gel filtration on large Sephadex® columns.
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