Ultrasensitive mouse enzyme linked immunosorbent assay

Body weight, postprandial blood glucose and insulin levels were determined immediately before sacrifice at all study points (4, 14 and 26 weeks) in db/db mice, as previously described [9] . Body weight, postprandial glucose and insulin levels were also determined in C57Bl6 mice after 18 weeks on HFD (26 weeks of age). Briefly, blood glucose levels were measured from nicked tails using the glucometer Optium Xceed (Abbott, United Kingdom). Blood for plasma insulin determination was collected from the tail vein into capillary tubes precoated with potassium-EDTA (Sarstedt, Nümbrecht, Germany). Plasma insulin levels were measured using ultrasensitive mouse enzyme-linked immunosorbent assay (ALPCO Diagnostics, Salem, NH).

Body weight, glucose and insulin levels were determined in at 30–32 weeks of age, before sacrifice [14 (link)]. Blood glucose levels were measured using the glucometer Optium Xceed (Abbott, UK) and plasma insulin samples were collected into tubes with potassium-EDTA (Sarstedt, Nümbrecht, Germany). Blood samples were centrifuged during 7 min, 6500 rpm at 4 °C, and plasma fraction was stored at −80 °C until processed. Plasma insulin levels were measured using ultrasensitive mouse enzyme-linked immunosorbent assay (ALPCO Diagnostics, Salem, NH, USA).

Body weight, postprandial blood glucose, and insulin levels were determined at 18 (before the commencement of the treatment) and 26 weeks of age as previously described [11 (link)]. Briefly, blood glucose levels were measured from nicked tails using the glucometer Optium Xceed (Abbott, UK). Blood for plasma insulin determination was collected from the tail vein into capillary tubes precoated with potassium-EDTA (Sarstedt, Nümbrecht, Germany). Blood samples were centrifuged during 10 min, 6500 rpm at 4 °C, and plasma fraction was stored at −80 °C until processed. Plasma insulin levels were measured using ultrasensitive mouse enzyme-linked immunosorbent assay (ALPCO Diagnostics, Salem, NH, USA).