The titrations to determine the kinetic constants of complex formation were designed with 10 different analyte concentrations, plus one duplicate concentration.
Concentrations for all markers determined via the Luminex100® system were obtained in duplicate; only concentrations with a coefficient of variation ≤ 0.5 were included.
All samples and standards were measured in duplicate and concentrations were determined from a standard curve using mean optical density values.
Finally, the Dietary Exposure Potential Model (DEPM) was used with the Dietary Checklist data to predict duplicate diet concentrations within our sample.
Serum samples were assayed in duplicate for concentrations of leptin, IL-1β, IL-6, MCP-1, and TNF-α using a multiplex cytokine assay (Millipore Milliplex, cat no. RCYTOMAG-80 K; Billerica, MA, USA) on a MAGPIX instrument (Luminex Technologies; Luminex, Austin, TX, USA) according to the manufacturer's instructions.
Ethylene produced by the bacteria was quantified using standard ethylene (C2H4, Product Number: 00489, Sigma-Aldrich) curve prepared in duplicates in concentrations ranging from 1 1000 nmol.
Ethylene produced by the bacteria was quantified using standard ethylene (C2H4, Product Number: 00489, Sigma-Aldrich) curve prepared in duplicates in concentrations ranging from 1 to 1000 nmol.
The same volume of anti-GST 5′ and 3′ SH2-PLA probes were gently mixed, and 2 μl of mixed assay probes were added to 2 μl of serially diluted GST protein in duplicate (GST concentration: 0.13 nM – 4.17 nM).
Sperm percentages of A + B grades of motility, that is, progressive motility, were assessed at 400× magnification on a microscope heating stage (37°C) in duplicate, and concentration was measured using a Bürker-Türk chamber at phase contrast (400×).
