Inhibin b gen 2 elisa

Aliquots of the culture media from all the experimental groups were collected and stored at −20°C for the assessment of AMH (AMH Gen II ELISA, Beckman Coulter; intra-assay CV = 3.89%; inter-assay CV = 5.77%) and inhibin B (Inhibin B Gen II ELISA, Beckman Coulter, Webster, TX, USA; intra-assay CV = 2.81%; inter-assay CV = 4.33%) secretion as previously described (50 (link)).

Aliquots of culture media from all the experimental groups were collected and stored at -20°C for subsequent assessment of AMH (AMH Gen II ELISA, Beckman Coulter, Webster, TX, USA) and inhibin B (inhibin B Gen II ELISA, Beckman Coulter) secretion levels as previously described (34 (link)).

At the time of delivery, a blood sample of 10–30mL was collected from the umbilical cord and stored at -80°C for later analysis.
The following hormone levels in 294 stored cord blood samples (135 boys and 159 girls) were measured. Testosterone (T), estradiol (E), and progesterone (P) levels were measured using LC-MS/MS [16 (link), 17 (link)]. An immunoradiometric assay was used to measure luteinizing hormone (LH) (Spac-S LH Kit, TFB, Inc., Tokyo Japan) and follicle-stimulating hormone (FSH) levels (Spac-S FSH Kit, TFB, Inc., Tokyo Japan). Inhibin B levels were measured using an enzyme-linked immunosorbent assay (Inhibin B Gen II ELISA, Beckman Coulter, Inc., CA, USA). An enzyme immunoassay (Insulin-like 3 (INSL3) / RLF (Human)—EIA Kit, Phoenix Pharmaceuticals, Inc. CA, USA) was used to measure INSL3 levels. INSL3 was measured in males because it reflects Leydig cell function. It was also measured in 20 randomly selected samples from females. All sex hormone measurements were performed by Aska Pharma Medical Co., Ltd. (Kanagawa, Japan).

Aliquots of the culture media from all the experimental groups were collected and stored at −20 °C for the assessment of AMH (AMH Gen II ELISA, Beckman Coulter; intraassay CV = 3.89%; interassay CV = 5.77%) and inhibin B (inhibin B Gen II ELISA, Beckman Coulter, Webster, TX, USA; intraassay CV = 2.81%; interassay CV = 4.33%) secretion, as previously described [7 (link)].

Serum hormone concentrations were determined by enzyme immunoassay using commercially available kits “Steroid IFA-Testosterone-01”, “Gonadotropin IFA-LH”, “Gonadotropin IFA-FSH” (Alkor Bio, St. Petersburg, Russia), “Estradiol-IFA” (Xema Medica, Moscow, Russia) and “Inhibin B Gen II ELISA” (Beckman Coulter, Pasadena, CA, USA). The ranges of evaluated concentrations for total testosterone (T), estradiol (E₂), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and inhibin B (InhB) were 0.2–50 nmol/L, 0.1–20 nmol/L, 2.0–100 mME/mL, 20–90 mME/mL, and 12–105 pg/mL, respectively. The sensitivities for T, E₂, FSH, LH, InhB were 0.2 mmol/L, 0.025 nmol/L, 0.25 mME/mL, 0.25 mME/mL, 2.6 pg/mL, respectively. The intra- and interassay coefficients of variation were as follows: for T < 8.0%; E₂ < 8.0%; FSH < 8.0%; LH < 8.0%, InhB ≤ 6.8%.

Aliquots of culture media from untreated and cisplatin plus EPA treated SCs were collected and stored at -20°C for subsequent assessment of AMH (AMH Gen II ELISA, Beckman Coulter, Webster, TX, USA) and inhibin B (inhibin B Gen II ELISA, Beckman Coulter) secretion levels as previously described (54 (link)).

Serum inhibin B was determined using an enzyme-linked immunoassay specific for human inhibin B (Inhibin B Gen II ELISA^®, Immunotech Beckman-Coulter Co., Prague, Czech Republic). Intraassay coefficients of variation were 14% for a serum inhibin B concentration of 111 pg/mL, and 9.8% for 479 pg/mL. Inter-assay coefficients of variation were 14% for a serum inhibin B concentration of 12 pg/mL, and 7.7% for 210 pg/mL. When serum inhibin B levels was undetectable, a value of 7.2 pg/mL, corresponding to the limit of quantification (functional sensitivity), was attributed.