Quantikine elisa mouse adiponectin acrp30 immunoassay

Quantification by ELISA of intra- and extracellular adiponectin levels in 3T3-L1 cells expressing GFP-Rab34 or silenced for this protein by Rab34 siRNA treatment was carried out using Quantikine ELISA Mouse Adiponectin/Acrp30 Immunoassay (R&D Systems Europe, Abingdon, UK) using manufacturer’s instructions.

Plasma glucose, insulin, adiponectin, and leptin levels were measured by Autokit Glucose (Wako, Osaka, Japan), Mouse Insulin ELISA KIT (U‐type) (Shibayagi, Japan), Quantikine^® ELISA Mouse Adiponectin/Acrp30 Immunoassay (R&D Systems, Minneapolis, MN, USA), and Quantikine^® ELISA Mouse/Rat Leptin Immunoassay (R&D Systems), respectively. t‐Cho, TG, and NEFA were measured with LabAssay^™ Cholesterol (Wako), LabAssay^™ Triglyceride (Wako), and LabAssay^™ NEFA (Wako), respectively. All assays were performed according to the manufacturers' protocols. Plasma 3‐HB levels were measured by modification of a previously reported method (Hansen & Freier, 1978). Briefly, plasma was added to a reaction buffer, containing 80 mm Tris–HCl (pH 9.5) and 4 mm β‐NAD⁺, and incubated at 37 °C for 5 min. Reactions were initiated by addition of 0.37 U mL⁻¹ 3‐hydroxybutyrate dehydrogenase. Changes in absorbance were measured for 10 min at 340 nm using a SpectraMax Plus384 (Molecular Devices, Sunnyvale, CA, USA).