Blood glucose was measured using Touch Sure Step strips (Life Scan, Johnson and Johnson, Milpitas, CA, USA). Total cholesterol, triglyceride (TG), and non-esterified fatty acid (NEFA) levels were measured with commercially available kits (Cholesterol-CH7945; TG-TR313 and NEFA-FA115, Randox Laboratories, Antrim, UK). Plasma insulin levels were measured with an ELISA kit (Mercodia 10-1250-01; Uppsala, Sweden).
Nefa fa115
Manufactured by Randox
Sourced in United Kingdom
The NEFA-FA115 is a laboratory equipment designed to measure non-esterified fatty acids (NEFAs) in various samples. It utilizes an enzymatic colorimetric method to quantify NEFA levels accurately. The core function of this product is to provide reliable and precise NEFA analysis for research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Elisa kit, by Mercodia (1 mentions)
Urea, by DiaSys (1 mentions)
Cholesterol, by DiaSys (1 mentions)
Gl 32k, by Merck Group (1 mentions)
Vacuette, by Greiner (1 mentions)
Cobas mira, by Roche (1 mentions)
Gluc pap gl364, by Randox (1 mentions)
Ranbut rb1007, by Randox (1 mentions)
Pentra 400 analyzer, by Horiba (1 mentions)
Nefa hr 434 91795, by Fujifilm (1 mentions)
Mouse insulin elisa, by Mercodia (1 mentions)
Gluc3, by Roche (1 mentions)
Vitros 5.1 fs, by Ortho Clinical Diagnostics (1 mentions)
Vacutainer vacuum system, by BD (1 mentions)
Ranbut, by Randox (1 mentions)
Cobas bio, by Roche (1 mentions)
Cobas c701 702, by Roche (1 mentions)
4 protocols using nefa fa115
1
Metabolic Biomarker Quantification Protocol
2
Plasma Metabolite and Hormone Profiling
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After morning milking and before feeding, blood samples were drawn daily from one jugular vein us-ing evacuated tubes coated with EDTA for plasma preparation, and with clot activator for serum harvest, respectively (Vacuette, 9 mL with K 3 EDTA, cat. no. 455036; Vacuette, 9 mL with serum clot activator, cat. no. 455092; Greiner Bio-One International GmbH). Samples in EDTA tubes were put immediately on wet ice, whereas serum tubes were kept at ambient temperature for around 2 h until centrifugation. After centrifugation at 2,000 × g (+4°C, 20 min), aliquots of 1.5 mL were frozen at -80°C until further analyses. In plasma, various metabolites were measured using commercially available enzymatic kits with an autoanalyzer (Cobas Mira, Roche): glucose (GLUC-PAP GL364), NEFA (NEFA FA115), and BHB (Ranbut RB1007) kits were obtained from Randox Laboratories Ltd., and kits for measurements of Urea (Urea FS 1.3101 99 10 021), total Cholesterol (Cholesterol FS 1.1350 99 10 021), and triglycerides (Triglycerides FS 1.5760 99 10 021) from DiaSys Diagnostic Systems GmbH.
Glucagon in plasma was measured by a commercially available RIA (catalog no. GL-32 K, EMD Millipore), whereas concentrations of insulin and IGF-1 were measured by RIA as described earlier by Vicari et al. (2008) (link).
Glucagon in plasma was measured by a commercially available RIA (catalog no. GL-32 K, EMD Millipore), whereas concentrations of insulin and IGF-1 were measured by RIA as described earlier by Vicari et al. (2008) (link).
3
Measuring Murine Glucose Metabolism and Lipid Profile
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Murine blood glucose measurements were performed after a 2-h fasting period using the tail-tip method to collect sufficient blood samples. The effects of the interventions on blood glucose levels were determined by the Optium EZ monitoring system (Abbott, Lake Bluff, IL, USA) and glucose tolerance testing (IPGTT), as previously described.29 (link) Briefly, ALX-induced or NOD/ShiLtJ mice that underwent overnight fasting for 16 h were administered an intraperitoneal injection of glucose (2 g/kg body weight). Blood glucose levels were measured at 10-, 15-, 30- and 120-min timepoints. Serum triglycerides and total cholesterol were measured spectrophotometrically using an enzymatic assay kit (ABX Pentra A11A01640, Horiba-ABX, Montpellier, FRA). Serum free fatty acids were measured by the acyl-CoA synthase and acyl-CoA oxidase methods (NEFA-HR 434-91795, Wako Chemicals, Richmond, VA, USA). Serum β-hydroxybutyrate was quantified by an enzymatic assay (NEFA FA115, Randox Lab, Crumlin, UK). The biochemical parameters were determined using a Pentra 400 analyzer (Horiba-ABX, Montpellier, FRA) following the manufacturer’s instructions. Plasma insulin content was measured in duplicate without dilution by using a standard fluorescence direct sandwich ELISA system (catalog no. 10-1247-01, Mouse Insulin ELISA, Mercodia Inc, NC, USA) according to the manufacturer’s protocol.
4
Fasting Blood Metabolic Profiles
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Blood samples were collected in fasting state from the antecubital vein (BD Vacutainer® vacuum system, Franklin Lakes, NJ, USA), after approximately 8 h of sleep (last meal no more than 2 h before bedtime). The following were determined in the blood: glucose concentration (GLU), ketone bodies (KB) (β-hydroxybutyrate—BHB), triglycerides (TG) and free fatty acids (FFA). GLU concentration was determined in the plasma (EDTA and glycolysis inhibitors: sodium fluoride and potassium oxalate). Triglycerides (TG), FFA and BHB, were determined in the serum (clotting activator). The assays were carried out using commercial reagent kits according to the procedure indicated by the manufacturers.
The detection range was, respectively: 0.11–41.6 mmol/L for glucose (GLUC3 Roche Diagnostics International Ltd., Germany), 0.07–2.24 mmol/L for FFA (NEFA FA 115, Randox Laboratories Ltd., UK), 0.1–5.75 mmol/L for 3-hydroxybutyrate (Ranbut, Randox Laboratories Ltd., UK) and 0.11–5.93 mmol/L for triglycerides (TRIG, Ortho-Clinical Diagnostics, France). The determinations were performed using the Cobas c 701/702 (glucose) and Cobas Bio (FFA, BHB) analysers by Roche Diagnostics International Ltd. (Germany) and Vitros 5.1 FS (TG) by Ortho-Clinical Diagnostics (France). The intra-assay % coefficients of variation were: KB 5.2%, TG 1.1%, FFA 4.3%, GLU 1.1%.
The detection range was, respectively: 0.11–41.6 mmol/L for glucose (GLUC3 Roche Diagnostics International Ltd., Germany), 0.07–2.24 mmol/L for FFA (NEFA FA 115, Randox Laboratories Ltd., UK), 0.1–5.75 mmol/L for 3-hydroxybutyrate (Ranbut, Randox Laboratories Ltd., UK) and 0.11–5.93 mmol/L for triglycerides (TRIG, Ortho-Clinical Diagnostics, France). The determinations were performed using the Cobas c 701/702 (glucose) and Cobas Bio (FFA, BHB) analysers by Roche Diagnostics International Ltd. (Germany) and Vitros 5.1 FS (TG) by Ortho-Clinical Diagnostics (France). The intra-assay % coefficients of variation were: KB 5.2%, TG 1.1%, FFA 4.3%, GLU 1.1%.
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