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The liquid phase, double antibody radioimmunoassays for rat LH and FSH were studied with respect to the minimum duration required for their completion using NIAMDD (National Institute of Arthritis, Metabolism and Digestive Diseases) reagents at their recommended dilutions. The minimum assay duration, which still allows maximum sensitivity, at temperatures not exceeding room temperature is 3 days for the LH assay and 4 days for the FSH assay. The extra day for the FSH assay is due to apparently nonreversible binding of rat FSH to anti-rat FSH. Such nonequilibrium binding was also observed under certain conditions with the LH assay. The greatest tracer binding was observed at a I-Ab (rabbit anti-LH or anti-FSH) incubation of 1 day at 27 degrees C plus 1 day at 4 degrees C followed by 1 additional day at 4 degrees C with the II-Ab (antirabbit gamma-globulin).
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Findley et al. (1982) studied this question.
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