Amh gen 2 elisa assay kit

Blood samples obtained by venipuncture were centrifuged, within 30 minutes from the collection, for 10 min at 3500 rpm, 4°C. Aliquots of each serum were frozen at −80°C and stored for subsequent assays of AMH, FSH and E2. To measure serum AMH levels, AMH-Gen II ELISA assay kit (Beckman Coulter) was used. The lowest detection limit of AMH is 0.08 ng/mL and the intra and interassay coefficients of variation below 3.4% and 4.0% respectively. The ECLIA method used to measure the levels of serum FSH and E2 by COBAS e411 auto-analyser (Roche Diagnostics). The lowest detection limit for FSH is 0.100 IU/L, with the intra- and interassay coefficients of variation below 2.6% and 3.5% respectively. The lowest detection limit for E2 is 18.4 pmol/L, with the intra- and interassay coefficients of variation of 2% and 3%, respectively.
Giving the poor reliability of the FSH value in presence of Estradiol levels greater than 70 pg/mL, women with basal E2 levels greater than this cut off were asked to come back at the beginning of the subsequent menstrual cycle, and their data were excluded from the analysis.

Serum anti-Müllerian hormone (AMH) concentration was determined using the commercially available AMH Gen II ELISA assay kit (Beckman Coulter, California, A79766) according to the manufacturer's instructions. AMH levels were measured for serum samples of 12-month-old WT (n = 6), Asc^−/− (n = 6), and Nlrp3^−/− mice (n = 6). Serum samples were assayed in duplicates and the absorbance was measured using the CLARIOstar microplate reader (BMG Labtechset, Ortenberg, Germany).