Bio plex pro human diabetes adipsin and adiponectin assays

Plasma adiponectin concentration was assessed by flow cytometry on a double beam laser automated analyser (Bio-Plex Protein Assay System, Bio-Rad, USA) using commercial test kits (Bio-PlexPro Human Diabetes Adipsin and Adiponectin Assays, Bio-Rad, USA), according to the manufacturer's protocol.

Adiponectin and adipsin were measured using Bio-Plex Pro Human Diabetes Adipsin and Adiponectin Assays (Bio-Rad Laboratories Ltd, Hertfordshire, U.K, Catalog number #171A7002M). Plasma samples were diluted 380 times with sample dilution buffer for adiponectin and adipsin measurement as per the manufacturer’s instructions. Coefficient of variation and minimal detection levels for adiponectin was CV < 7.8% and minimal detection level 6.9 pg/ml and for adipsin was CV <11.8% and minimal detection level 9.3 pg/ml. Five-parameter logistic regression algorithms built into the Bio-Plex Manager 6.0 software were used to assess levels of adiponectin and adipsin with reference to standards. Analysis was conducted using a Bioplex-200 instrument according to the manufacturer’s instructions (BIO-RAD, Hertfordshire, UK).

The serum levels of pancreatic hormones (insulin and glucagon), incretins (ghrelin, GIP and GLP-1) and adipokines (adiponectin, leptin, resistin, adipsin, PAI-1, visfatin) were estimated by Bio-Plex Pro Human Diabetes 10-Plex Assay and Bio-Plex Pro Human Diabetes Adipsin and Adiponectin Assays, as per the kit protocol (Biorad, USA). The lowest detection limits for the tested cytokines were: Insulin- 1.7 pg/ml, Glucagon- 15.7 pg/ml, Ghrelin- 8.01 pg/ml, GIP- 3.41 pg/ml; GLP-1- 5.91 pg/ml, Adiponectin- 160 pg/ml, Leptin- 11.5 pg/ml, Resistin-2.3pg/ml, Adipsin-43pg/ml, PAI-1- 14.24 pg/ml; Visfatin- 51.3 pg/ml.