Human adiponectin elisa kit

Body composition was measured by air-displacement plethysmography using the Bod Pod Gold Standard system (Bod Pod 2007 A, Life Measurement, Concord, CA, software versions 4.2.0 and 5.2.0.) using gestational-age specific equations according to our previously described protocol^{29 (link)}. Leptin and adiponectin were analysed at the Clinical Chemistry Laboratory, Sahlgrenska University Hospital (accredited in accordance with the International Standard ISO 15189:2007). Leptin (Human Leptin Quantikine, R&D Systems, Minneapolis, MN; interassay coefficient of variation, 8.0% at 9 μg/l), the soluble leptin receptor (sOB-R) (Human Leptin R Quantikine, R&D Systems) and adiponectin (Human Adiponectin ELISA kit, Millipore, Billerica, MA; interassay coefficient of variation, 7.0% at 10.5 mg/l) were analysed using ELISA. sOB-R measurements were conducted for a subset of women, i.e. the women that were included in the full PONCH study of both mother and child²¹ ; this resulted in 84 NW women and 33 OB women that had complete sOB-R measurements in both T1 and T3.

HbA1c was analysed immediately with a point-of-care analyser (Afinion AS100; Axis-Shield, Oslo, Norway). Glucose, insulin, cholesterol (total, low-density lipoprotein (LDL), high-density lipoprotein (HDL)), triglycerides, leptin, adiponectin, C-reactive protein, and FFAs were analysed at the accredited Clinical Chemistry Laboratory, Sahlgrenska University Hospital (International Standard ISO 15189:2007). ELISA was used to measure adiponectin (Human Adiponectin ELISA kit, Millipore, Billerica, MA; interassay coefficient of variation, 7.0% at 10.5 mg/L) and leptin (Human Leptin Quantikine, R&D Systems; interassay coefficient of variation, 8.0% at 9 μg/L). All other assays were routinely done with a cobas modular system (Roche Diagnostics). Homeostatic model assessment of insulin resistance (HOMA-IR) was calculated as (fasting glucose × fasting insulin)/22.5 [13 (link)]. Adipose tissue insulin resistance (ADIPO-IR) was calculated as fasting FFA × fasting insulin [14 (link)]. Due to occasional technical problems either with blood sampling or blood analysis, full sets for analysis were not available for all parameters at all time points. See Table 1 for n numbers.

Adiponectin was measured using a latex-enhanced turbidimetric immunoassay (Lot#230222) (Denka Co. Ltd., Tokyo, Japan) on a Beckman Coulter AU680 analyzer (Beckman Coulter, Krefeld, Germany). The coefficients of variation were 2.0% and 0.9% at levels of 2.0 and 7.7 µg/mL, respectively. The latex assay was compared with a human adiponectin ELISA kit (Lot#3940441) (Millipore via Sigma-Aldrich, Vienna, Austria) by running in parallel 51 sera with a wide range of adiponectin concentrations. Both assays resulted in comparable values and correlated very well. The acceptance criteria were slope 1.0 ± 0.1 and R ≥ 0.9.