Local Release of Steroid Hormones: MATERIALS AND METHODS(5)

The EIA for E2 was carried out as described previously. The standard curve ranged from 2 to 2000 pg/ml, and the ED50 of the assay was 120 pg/ml. The intra- and interassay CVs were 6.1% and 8.9%, respectively.

The A EIA was identical to the P4 EIA. Basically, standards or samples were incubated with 100 ^l polyclonal antibody (raised in a rabbit against A-3-CMO-BSA; Cosmo Bio Co., Tokyo, Japan) solution (1:500 000) and 100 ^l A-3-CMO-horseradish peroxidase (1:70 000; Cosmo Bio Co.) for 16 h at 4°C. The standard curve ranged from 2 to 1000 pg/ml, and the ED50 of the assay was 130 pg/ml. The intra-and interassay CVs were 6.8% and 8.1%, respectively. The cross-reactivities of the antibody were 100% for A, 35% for 5a-androstenedione, 4.5% for dehydroepiandrosterone (DHA), 1.5% for A, 1.2% for P4, 6.0% for testosterone, 0.3% for cortisol, 0.2% for corticosterone, and 0.01% for estradiol. buy prednisone

The EIA for PGE2 is described elsewhere. The standard curve ranged from 30 to 14 200 pg/ml, and the ED50 of the assay was 350 pg/ml. The intra- and interassay CVs were 9.5% and 12.5%, respectively. The cross-reactivities of the antibody were 100% for PGE2, 60% for PGB2, 15% for PGEb 4% for PGA2, 3% for PGF2a, 0.1% for PGD2, and 0.04% for 15-keto-PGE2.

This entry was posted in Steroid Hormones and tagged Bovine Mature Follicles, Endothelin, Luteinizing Hormone, Prostaglandin, Steroid Hormones.