Development and validation of a sensitive radioimmunoassay procedure for estimating FSH in mithun (Bos frontalis) plasma.

The present study was designed to develop and validate a simple and sensitive radioimmunoassay (RIA) procedure to estimate FSH in mithun (Bosfrontalis) plasma. The assay was carried out in 100 [L of mithun plasma. The bovine FSH standards (10 to 5000 pg/100 microL/tube) in hormone-free plasma were used in the assay. The sensitivity of the assay was 20 pg/100 microL/tube, which corresponded to 0.20 ng/mL plasma. The 50% relative binding sensitivity of the assay was 80 pg/100 microL/tube, which corresponded to 0.80 ng/mL plasma. The intra- and inter-assay coefficients of variation were 4.6% and 12.4%, respectively. The biological validation of the assay was carried out in plasma samples that were collected during different stages of the estrous cycle. In the entire estrous cycle, plasma FSH concentration (p < 0.01) attained two peaks (on day 3 to 4 before estrus 5.1 +/- 0.3 ng/mL and on the day of estrus 6.9 +/- 0.2 ng/mL). FSH concentration remained at basal level (1.3 +/- 0.1 to 1.6 +/- 0.2 ng/mL) during day 4 to 16 of the estrous cycle. The concentration of plasma FSH was found to be significantly (p < 0.05) higher (4.9 +/- 0.3 to 6.8 +/- 0.5 ng/mL) until 48 h following the estrus onset. In conclusion, the RIA procedure that was developed in the current study is sufficiently reliable and sensitive to estimate different physiological levels of FSH in mithun plasma.