Enzyme linked immunosorbent assay elisa kits

Baseline venous blood samples were collected after overnight fasting at 3-month intervals before conception. After collecting baseline samples, participants received different doses of folic acid (5 mg/d vs. 0.5 mg/d) until 36 wk of pregnancy. The participants were contacted every week and were asked during each visit to declare whether they took folic acid supplementation or not. Second sampling was done in the 36 wk of pregnancy. The participants that had an abortion, multiple pregnancies, usage of calcium and ferrous sulfate during pregnancy and did not complete the follow-up were excluded from the study. Serums were collected after centrifugation (at 3000 RPM) of the samples for 20 min, and then stored at −70 °C until analysis. We used the date of the mother’s last menstrual period (LMP) for calculating Gestation age. Serum OPG and sRANKL concentrations were measured by commercially Enzyme-Linked Immunosorbent Assay (ELISA) kits (Bioassay Technology Laboratory, China, with intra- and inter- assay CV of <10% and <12%, respectively). TNFα was analyzed in serum by ELISA using Immunodiagnostic kit (DIAsource Immuno Assay S.A, Belgium, with intra- and inter- assay CV of 6.6% and 4.5%, respectively).

Metformin (hydrochloride) and AMX were purchased from Sigma-Aldrich (Germany). The following monoclonal antibodies (mAbs) were used for flow cytometric analysis: anti-CD4 FITC, anti-Foxp3 PE, anti-CD25 PerCP, anti-RORγt PE, and Mouse Regulatory T cell Staining Kit, purchased from BD Biosciences (Germany). Enzyme-linked immunosorbent assay (ELISA) kits were purchased from Bioassay Technology Laboratory (China). The DNA extraction kit, viral RNA kit, and RNeasy mini kit were obtained from Qiagen (Germany). cDNA synthesis kit/reagents were purchased from Dena Zist Asia (Iran).