Blood specimens were acquired by puncturing the retro-orbital plexus under isoflurane anesthesia. Serum triglycerides and total cholesterol were measured by means of a dry chemistry analyzer (Fuji DRI-CHEM NX500i, Fujifilm, Wiesbaden, Germany). LDL cholesterol was analyzed by using an enzymatic assay (Crystal Chem, Zaandam, Germany). The serum concentration of leptin (Merck Millipore, Darmstadt, Germany), insulin (Crystal Chem), free thyroxine (fT4; Cusabio, Houston, TX, USA), ET-1 (R&D systems, Bio-Techne, Wiesbaden, Germany), and FGF21 (BioVendor, Karasek, Czech Republic) were determined by ELISA kits according to the provided instructions.
Enzymatic assay
Manufactured by Crystal Chem
Sourced in Germany
The enzymatic assay is a laboratory tool used to measure the activity or concentration of a specific enzyme in a sample. It provides a quantitative analysis of the enzyme's presence and function, without interpretation or extrapolation beyond its core purpose.
Automatically generated - may contain errors
2 protocols using enzymatic assay
1
Comprehensive Metabolic Profiling Protocol
2
Intraperitoneal Glucose Tolerance Test
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An intraperitoneal glucose tolerance test (IPGTT) was performed before and after the treatment with EET-A following the National Mouse Metabolic Phenotyping Centers glucose tolerance test protocols (2) . Mice fasted 5 h received an i.p. injection of 20% w/v glucose (final 2 g/kg body wt), and tail-vein blood glucose was measured at baseline, 15, 30, 45, 60, and 90 min. The area under the curve for glucose was calculated using the trapezoidal method.
Plasma samples were collected from 5-h fasted mice for glucose and insulin assay. Glucose was analyzed by enzymatic assay (CrystalChem, Elk Grove Village, IL).
Plasma insulin was determined by radioimmunoassay by the Vanderbilt Mouse Metabolic Phenotyping Center Analytical Core Laboratory. The updated HOMA of insulin resistance (HOMA2-IR) was calculated using fasting glucose and insulin (HOMA2 calculator v2.2, Oxford, U.K.) (13) .
Plasma samples were collected from 5-h fasted mice for glucose and insulin assay. Glucose was analyzed by enzymatic assay (CrystalChem, Elk Grove Village, IL).
Plasma insulin was determined by radioimmunoassay by the Vanderbilt Mouse Metabolic Phenotyping Center Analytical Core Laboratory. The updated HOMA of insulin resistance (HOMA2-IR) was calculated using fasting glucose and insulin (HOMA2 calculator v2.2, Oxford, U.K.) (13) .
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