Serum was separated by centrifugation (3000 rpm, for 8 min) at room temperature and stored at −20°C for the serum analysis. The glucose (monoreagent-K082), triglyceride (monoreagent–K117) and total cholesterol (monoreagent-K083) concentrations were measured by a colorimetric assay (Bioclin Systems II®, Quisaba, Bioclin, Belo Horizonte, MG, Brazil). The serum analyses for insulin, leptin and testosterone were performed using the following commercially available enzyme-linked immunosorbent assay (ELISA) kits: rat/mouse insulin kit (Millipore-Cat. EZRMI-13 k – St Charles, MO, USA), rat leptin kit (Millipore-Cat. EZRL-83K, St Charles, MO, USA) and general testosterone kit (Uscn-Cat. E90458Ge – Wuhan, China). All samples were analyzed in duplicate with an intra-assay coefficient of variation of 1.4%.
Rat mouse insulin kit
Manufactured by Merck Group
Sourced in United States
The Rat/Mouse Insulin kit is a laboratory product designed to measure insulin levels in rat and mouse samples. It is a quantitative assay that employs the principles of a sandwich enzyme-linked immunosorbent assay (ELISA) technique. The kit includes the necessary reagents and materials required to perform the analysis.
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Lab products found in correlation
7 protocols using rat mouse insulin kit
1
Serum Biochemistry and Hormone Analysis
2
Serum Biomarker Profiling Protocol
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After the blood was collected, the serum was separated by centrifugation at room temperature (3000 rpm, 8 min) and stored at −20°C. Glucose, total cholesterol and triglyceride (TG) concentrations were measured using a colorimetric assay (Bioclin; Belo Horizonte, Minas Gerais, Brazil). An automatic spectrophotometer was used following the instructions recommended by the manufacturer of Bioclin commercial kits: glucose monoreagent-K082, cholesterol monoreagent-K083 and triglycerides monoreagent-K117. Serum analyses for insulin (Rat/Mouse Insulin kit, Millipore - Cat. EZRMI-13 k – St Charles, Missouri, USA) and testosterone (General Testosterone kit, Uscn - Cat. E90458Ge – Wuhan, China) were performed using commercially available enzyme-linked immunosorbent assay (ELISA) kits.
IR was calculated using the HOMA-IR (homeostasis model assessment for IR index): insulin*glucose/22.5.
IR was calculated using the HOMA-IR (homeostasis model assessment for IR index): insulin*glucose/22.5.
3
Evaluating Metabolic Biomarkers in Rodents
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To evaluate triglycerides (TAG) and cholesterol (CHOL) levels, blood samples were collected on the experimental days after overnight fasting. By enzymatic colorimetry, serum aliquots were used to measure the levels of TAG and cholesterol.
To evaluate fasting glucose, blood samples were collected after overnight fasting by decapitation or from the tail. Glycaemia was determined on an Accu-Chek Performa glucometer.
To determine serum insulin and leptin, blood samples were collected on the experimental days after overnight fasting. Serum insulin were determined using Rat/Mouse Insulin Kit from Millipore and serum leptin were determined using Mouse Leptin ELISA kit.
Hepatic total lipids content was evaluated as proposed by Folch and colleagues [35 (link)].
To evaluate fasting glucose, blood samples were collected after overnight fasting by decapitation or from the tail. Glycaemia was determined on an Accu-Chek Performa glucometer.
To determine serum insulin and leptin, blood samples were collected on the experimental days after overnight fasting. Serum insulin were determined using Rat/Mouse Insulin Kit from Millipore and serum leptin were determined using Mouse Leptin ELISA kit.
Hepatic total lipids content was evaluated as proposed by Folch and colleagues [35 (link)].
4
Plasma Glucose and Insulin Measurement
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Plasma glucose was determined using Accuchek glucometer (Roche. Germany). Plasma insulin was measured with an ELISA insulin assay (Rat/Mouse Insulin kit, Millipore).
5
Serum Lipid and Insulin Analysis
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Serum was obtained after blood centrifugation (1000×g for 10 min at 4 °C). Total serum cholesterol, high-density lipoprotein (HDL), very low-density lipoprotein (VLDL), lowdensity lipoprotein (LDL) and triglycerides were assayed using enzymatic kits (Wiener ® , Argentina). Serum insulin was measured by chemiluminescence using a Rat/Mouse Insulin Kit (Millipore ® , Billerica, USA) and ADVIA-Centaur equipment.
6
Blood Lipid and Metabolic Biomarker Analysis
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Serum concentrations of free fatty acids (FFAs), total cholesterol (TC), low density lipoprotein (LDL)-c, high density lipoprotein (HDL)-c and triglycerides (TGs) were analyzed using commercial kits (Randox Laboratories Ltd., Crumlin, UK) in accordance with the manufacturer's protocol, and a Hitachi 8060 automatic biochemical analyzer (Hitachi Co., Ltd, Tokyo, Japan). TC and TG levels were determined by CHOD-PAP and GPO-PAP colorimetric end-point assays, respectively, using Randox Total Cholesterol (cat. no., CH200) and Triglycerides (cat. no., TR1697) kits. FFAs were measured by the non-esterified fatty acids (NEFA) colorimetric method using a Randox NEFA kit (cat. no., FA115). HDL-c and LDL-c were quantified by a direct clearance method using Randox Direct HDL-c (cat. no., CH2652) and Direct LDL-c kits (cat. no., CH2656). Blood glucose was measured with a OneTouch® Ultra™ Blood Glucose Monitoring System. Serum insulin and adiponectin levels were determined using a rat/mouse insulin kit (Linco; EMD Millipore, Billerica, MA, USA) and adiponectin enzyme-linked immunosorbent assay kit (cat. no. MRP300; R&D Systems, Inc., Minneapolis, MN, USA) in accordance with the manufacturer's protocol.
7
Quantifying Cytokine Levels via ELISA
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The enzyme-linked immunosorbent assay (ELISA) was performed with kits with serum samples from patients and mice according to the manufacturer's protocol. The human-IL6 kit (EHC007), mouse JE/MCP1/CCL2 (EMC113) kit, mouse IL-6 ELISA kit (EMC004), and mouse KC/IL-8/CXCL1 ELISA kit (EMC104) were purchased from NeoBioscience Technology Company (ShenZhen, China). The rat/mouse insulin kit (EZRMI-13K) was purchased from EMD Millipore Corporation. A mouse MMP12 ELISA kit (ARG81803) was purchased from Arigobiolaboratories company. The human CXCL1/KC kit was purchased from Multi Science Company. The data of ELISA was analyzed by using Curve Expert1.4 software.
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