Book contents
- Frontmatter
- Contents
- Introduction
- 1 Homeostasis: a fundamental organising paradigm in ecophysiology
- 2 Stress: the concept and the reality
- 3 Basic methods used in ecophysiological studies
- 4 Turnover methodology: theory and practice
- 5 Case studies of stress: incidence and intensity
- 6 Survival in deserts
- 7 Torpor and hibernation in cold climates
- 8 Marine birds and mammals
- 9 Conclusion
- Appendix 1 Population estimation methods
- Appendix 2 Estimation of food intake in Tiliqua rugosa
- Appendix 3 Simultaneous measurement of sodium and potassium concentration in plasma or urine using the IL 143 digital flame photometer
- Appendix 4 Determination of plasma urea nitrogen
- Appendix 5 Radioimmunoassay of testosterone in plasma
- Appendix 6 Preparation of ‘stripped plasma’
- Appendix 7 Radioimmunoassay of testosterone in faeces
- Appendix 8 The comparative method
- Appendix 9 Basic turnover equations
- References
- Index
Appendix 5 - Radioimmunoassay of testosterone in plasma
Published online by Cambridge University Press: 05 June 2012
- Frontmatter
- Contents
- Introduction
- 1 Homeostasis: a fundamental organising paradigm in ecophysiology
- 2 Stress: the concept and the reality
- 3 Basic methods used in ecophysiological studies
- 4 Turnover methodology: theory and practice
- 5 Case studies of stress: incidence and intensity
- 6 Survival in deserts
- 7 Torpor and hibernation in cold climates
- 8 Marine birds and mammals
- 9 Conclusion
- Appendix 1 Population estimation methods
- Appendix 2 Estimation of food intake in Tiliqua rugosa
- Appendix 3 Simultaneous measurement of sodium and potassium concentration in plasma or urine using the IL 143 digital flame photometer
- Appendix 4 Determination of plasma urea nitrogen
- Appendix 5 Radioimmunoassay of testosterone in plasma
- Appendix 6 Preparation of ‘stripped plasma’
- Appendix 7 Radioimmunoassay of testosterone in faeces
- Appendix 8 The comparative method
- Appendix 9 Basic turnover equations
- References
- Index
Summary
Extraction
Pipette 100 μl assay buffer into glass stoppered extraction tubes.
Pipette 10 μl plasma into each aliquot of buffer. Vortex briefly.
Pipette 2.5 ml diethyl ether into each sample, stopper, and vortex for 1 min. Allow to settle.
Using a Pasteur pipette, remove supernatant carefully to a clean assay tube.
Repeat, using a further 2.5 ml diethyl ether, and combine the supernatants.
Dry down under compressed air at 37 °C.
Assay
Prepare standards
Pipette duplicate 500 μl of each working standard into glass assay tubes.
Pipette duplicate 500 μl ethanol into tubes for 0 pg of steroid.
Dry down under compressed air at 37 °C.
Prepare assay and solvent blanks
Set up an assay tube as the assay blank. Antibody will be omitted from this tube, thus it is an estimate of any non-specific binding.
Pipette 5 ml diethyl ether into an assay tube as a solvent blank. Dry down.
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- Vertebrate EcophysiologyAn Introduction to its Principles and Applications, pp. 212 - 216Publisher: Cambridge University PressPrint publication year: 2003