Bio plex pro human diabetes panel

The Bio-Plex pro™ human diabetes panel (Bio-Rad Inc., Hercules, CA, USA) a Luminex-based magnetic bead assay, was used to quantify insulin, C-peptide, ghrelin, GIP, GLP-1, glucagon, leptin, resistin and visfatin and a separate Bio-Plex assay was used to quantify adiponectin and adipsin (due to different dilution factor) in plasma according to the manufacturer’s instructions. Each run included controls of known concentration for each cytokine and a blank.

Adipokine and hormonal biomarkers were measured in maternal serum samples using Bio-Plex Pro™ Human Diabetes Panel magnetic bead assays (Bio-Rad Laboratories, Hercules, CA, USA), in accordance with the manufacturer’s instructions with modifications as previously described [17 (link)]. The variables measured and their corresponding intra-assay CVs were C-peptide ~3%, ghrelin ~4%, gastric inhibitory polypeptide (GIP) ~6%, glucagon-like peptide-1 (GLP-1) ~4.5%, glucagon ~5%, leptin ~4%, plasminogen activator inhibitor-1 (PAI-1) ~3%, resistin ~4%, and visfatin ~5%.

Diabetes-related adipokines and hormones were determined in available first-thaw serum samples at baseline and at the end of each four-week intervention phase. These were measured using Bio-Plex Pro™ Human Diabetes Panel magnetic bead assays (Bio-Rad Laboratories, Hercules, CA, USA), according to the manufacturer’s instructions. Frozen serum samples were thawed on ice and 1:3 dilutions were prepared with the sample diluent provided in the kit. Diluted samples were processed using the manufacturer’s protocol with the following modifications: (i) beads were incubated with the detection antibodies for 45 min; and (ii) beads were incubated with streptavidin–phycoerythrin antibody for 20 min. The variables measured and their corresponding intra-assay CVs were C-peptide ~4%, ghrelin ~4%, gastric inhibitory polypeptide (GIP) ~3%, glucagon-like peptide-1 (GLP-1) ~4%, glucagon ~5%, leptin ~3%, plasminogen activator inhibitor-1 (PAI-1) ~2.5%, resistin ~5%, and visfatin ~4%.