Radioimmunoassay for the quantitative determination of Melatonin in biological samples.For research use only. In principle the Melatonin Research RIA kit will work for all kinds of biological samples. As a starting point it is recommended to perform a proof-of-principle (please contact our technical service for details). As a reference a proof of principle for rat plasma samples was performed using 100 µl of sample volume (please refer to chapter 8.). Melatonin - the major hormone secreted by the pineal gland - is a key modulator of annual and circadian biorhythms. Its circadian profile in body fluids is an excellent marker for the setting of the endogenous clock. Daytime plasma melatonin levels are low and rise in the evening (onset). Night-time levels peak at around 03.00 hrs. (acrophase) in most healthy humans. Onset, acrophase and offset have a stable phase relationship even when the phase of the melatonin profile is shifted. The assay procedure follows the basic principle of radioimmunoassay, involving competition between a radioactive and a non-radioactive antigen for a fixed number of antibody binding sites. The amount of 125I-labelled antigen bound to the antibody is inversely proportional to the analyte concentration of the sample. When the system is in equilibrium, the antibody bound radioactivity is precipitated with a second antibody in the presence of polyethylene glycol. The precipitate is counted in a gamma counter.Quantification of unknown samples is achieved by comparing their activity with a reference curve prepared with known standards. NAME: Melatonin Research RIADESCRIPTION: RIA for 100 determinations of Melatonin in biological samplesDESIGN: Common sample preparation. 7 standards, 2 controls, ready for use. Double antibody technique.SAMPLE VOLUME: 25 - 400 µlTOTAL ASSAY TIME: Sample preparation 1 hour and RIA 20-24 hoursSTANDARD RANGE: 0/ 30 - 10 000 pg/mlSENSITIVITY: 5.9 pg/ml (depending on sample volumel)STORAGE: 2 - 8 °CKIT: 100 DeterminationsADVANTAGES: Direct assayHigh specificity and sensitivity15 week shelf-lifeReady for use tracers
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