Rat insulin elisa

Manufactured by Mercodia

Sourced in Sweden, United States

The Rat Insulin ELISA is a laboratory equipment used for the quantitative determination of insulin levels in rat samples. It is a sandwich enzyme-linked immunosorbent assay (ELISA) designed to measure the concentration of insulin in biological samples.

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Close spelling variants of this product

Rat insulin ELISA kit (136 citations) Insulin (96 citations) Insulin ELISA (62 citations) Ultrasensitive Rat Insulin ELISA kit (39 citations) Ultrasensitive Rat Insulin ELISA (17 citations) High Range Rat Insulin ELISA kit (11 citations) ELISAs (9 citations) High Range Rat Insulin ELISA (7 citations) Ultrasensitive rat insulin ELISA system (4 citations) Rat insulin Elisa assay kit (4 citations)

54 protocols using rat insulin elisa

Serum Biomarkers in Fasted Rats

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Blood samples were taken from the heart of each animal in anesthetized rats. The rats were fasted overnight and supplemented with only tap water. The serum samples were sent to laboratory for analysis of glucose, insulin, nitrite, malondialdehyde (MDA), superoxide dismutase (SOD), and total antioxidant capacity (TAC).
The level of serum glucose was determined using quantitative diagnostic kits (Pars Azmoon, Iran). The level of insulin was measured using Mercodia Rat Insulin ELISA (Mercodia AB, Sylveniusgatan 8A, SE-754 Uppsala, Sweden). Mercodia Rat Insulin ELISA is a solid two-phase immunoassay. It is based on the direct sandwich technique, in which two monoclonal antibodies are directed against separate antigenic determinants on the insulin molecule. The level of nitrite in serum (stable nitric oxide metabolite) was measured using a colorimetric assay kit (ZelBio, Germany) that involves the Griess reaction. MDA levels of serum was quantified according to the manual methodology.[26 (link)27 (link)] Following the measurements, SOD activity and TAC in serum was measured using a colorimetric assay kit (ZelBio, Germany). In addition, for calculation of insulin resistance (IR), we measured the homeostatic model assessment (HOMA) index using the formula provided by Matthews et al.[11 (link)] Insulin (U/m) × (glucose [mmol/l]/22.5).

Ghofran O., Safari T, & Shahraki M.R. (2019). Effects of Eugenol on Pain Response to the Formalin Test and Plasma Antioxidant Activity in High Fructose Drinking Water in Male Rats. International Journal of Preventive Medicine, 10, 151.

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Streptozotocin-Induced Diabetic Rat Model

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Six-week-old male Slc:Wistar rats (Sankyo Labo Service Corporation, INC., Tokyo, Japan) were used (n = 36) in this study. All animal experiments were approved by the Institutional Animal Care, Animal Research and Use Committee of Tokyo Medical and Dental University (No. A2018-217A). Fig 1A shows a schematic representation of the study design. Diabetes (n = 18) was induced via intraperitoneal injection of STZ (60 mg/kg in saline) after 12 h of fasting [23 (link), 24 (link)]; rats in the control group (n = 18) were treated with the same volume of saline only. Body weights and plasma glucose levels were measured immediately before injection, 72 h after injection, and immediately before the surgical procedure. After 72 h, rats with fasting glucose levels above 350 mg/dL were considered diabetic rats [25 (link)]. Plasma insulin levels were determined using an enzyme-linked immunosorbent assay kit in accordance with the manufacturer’s instructions (Mercodia Rat Insulin ELISA, Mercodia, Oppland, Sweden).

Takeda K., Mizutani K., Matsuura T., Kido D., Mikami R., Noda M., Buranasin P., Sasaki Y, & Izumi Y. (2018). Periodontal regenerative effect of enamel matrix derivative in diabetes. PLoS ONE, 13(11), e0207201.

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Glycemic Biomarkers in Plasma

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Blood samples were collected in EDTA tubes and centrifuged at 5000 rpm for 10 min at 4 °C and plasma was kept at −20 °C until further analysis. Blood glucose was monitored by the Accu-Check^® Avia monitoring system (Roche Diagnostics, Rotkreuz, Switzerland). Plasma insulin was analysed according to the manufacturer’s instructions (Mercodia Rat Insulin ELISA, Mercodia AB, Uppsala, Sweden). HbA1c was measured from the tail vein and analysed using the DCA Vantage Analyzer (Siemens, Erlangen, Germany).

Sonne N., Larsen A.T., Karsdal M.A, & Henriksen K. (2022). The Impact of Exposure Profile on the Efficacy of Dual Amylin and Calcitonin Receptor Agonist Therapy. Biomedicines, 10(10), 2365.

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Plasma Insulin and Amino Acid Analysis

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In Experiment 2, plasma insulin levels were measured using a commercial ELISA kit for rat insulin (Mercodia Rat Insulin ELISA, Mercodia AB, Uppsala, Sweden). Plasma amino acids were measured by high-performance liquid chromatography with pre-column 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate derivatization [19 (link)].

Nakayama K., Tagawa R., Saito Y, & Sanbongi C. (2019). Effects of whey protein hydrolysate ingestion on post-exercise muscle protein synthesis compared with intact whey protein in rats. Nutrition & Metabolism, 16, 90.

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Glucose and Insulin Tolerance Assessments

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Glucose tolerance was tested by oral gavage of a 50% glucose solution to reach a glucose dosage of 2 g/kg body weight in 6-h fasted animals. Blood glucose response was measured on tail blood using Contour XT glucometer (Bayer). Insulin response was measured on plasma from tail blood collected in EDTA-coated tubes and stored on ice until centrifugation at 8400 × g for 6 min at 4 C. Insulin concentration was measured using Mercodia Rat Insulin ELISA (Mercodia AB) following the manufacturer’s protocol. Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) was calculated as HOMA-IR = (FPG + FPI)/135, where fasting plasma glucose (FPG) is in mmol/L and fasting plasma insulin (FPI) is in mU/L. The constant of 135 is based on an average value of 1 for healthy young animals^{104 (link)}. Insulin tolerance was tested by intraperitoneal injection of human insulin at a dosage of 0.75 UI/kg body weight (Actrapid, Novo Nordisk) in 4-h fasted animals. Blood glucose response was measured on tail blood using Contour XT glucometer (Bayer). Human insulin concentration was measured on tail blood (treated as above) using Mercodia Human Insulin ELISA (Mercodia AB) to ensure correct administration of insulin.

Peluso A.A., Lundgaard A.T., Babaei P., Mousovich-Neto F., Rocha A.L., Damgaard M.V., Bak E.G., Gnanasekaran T., Dollerup O.L., Trammell S.A., Nielsen T.S., Kern T., Abild C.B., Sulek K., Ma T., Gerhart-Hines Z., Gillum M.P., Arumugam M., Ørskov C., McCloskey D., Jessen N., Herrgård M.J., Mori M.A, & Treebak J.T. (2023). Oral supplementation of nicotinamide riboside alters intestinal microbial composition in rats and mice, but not humans. NPJ Aging, 9(1), 7.

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Blood Analysis of Glucose and Insulin

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Blood samples were collected in EDTA tubes and centrifuged at 5000 rpm for 10 minutes at 4°C, and plasma was kept at 220°C until further analysis. Blood glucose was monitored by Accu-Check Avia monitoring system (Roche Diagnostics, Rotkreuz, Switzerland). HbA1c levels were measured by DCA Vantage Analyzer (Siemens, Erlangen, Germany). Plasma levels of insulin (Mercodia Rat Insulin ELISA, RRID: AB_2811229; Mercodia AB, Uppsala, Sweden) and acetaminophen (Neogen Corporation's Acetaminophen ELISA Kit; Neogen Toxicology, KY) were analyzed according to manufacturer's instructions.

Larsen A.T., Sonne N., Andreassen K.V., Karsdal M.A, & Henriksen K. (2020). The Calcitonin Receptor Plays a Major Role in Glucose Regulation as a Function of Dual Amylin and Calcitonin Receptor Agonist Therapy. The Journal of pharmacology and experimental therapeutics, 374(1).

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Glucose Metabolism and Gastric Emptying

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Blood samples were collected in EDTA tubes and centrifuged at 5000 rpm for 10 min at 4 °C, and plasma was kept at −20 °C until further analysis. Blood glucose was monitored by an Accu-Check® Avia monitoring system (Roche Diagnostics, Rotkreuz, Switzerland). HbA1c levels were measured by a DCA Vantage Analyzer (Siemens, Erlangen, Germany). Plasma levels of insulin (Mercodia Rat Insulin ELISA, Mercodia AB, Uppsala, Sweden) were analyzed according to the manufacturers’ instructions. The gastric emptying rate was determined by measuring the acetaminophen in the plasma (Acetaminophen Forensic ELISA kit, Neogen Toxicology, KY, USA).

Andreassen K.V., Larsen A.T., Sonne N., Mohamed K.E., Karsdal M.A, & Henriksen K. (2021). KBP-066A, a long-acting dual amylin and calcitonin receptor agonist, induces weight loss and improves glycemic control in obese and diabetic rats. Molecular Metabolism, 53, 101282.

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Metabolic Biomarker Quantification Protocol

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Enzyme-linked immunosorbent assay (ELISA) kits were used to determine plasma insulin and leptin levels, specifically Mercodia Rat Insulin ELISA (Mercodia AB, Uppsala, Sweden) and Rat Leptin Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA). Circulating triglycerides (TG), non-esterified fatty acids (NEFA), and myo-inositol were measured by using the following enzymatic colorimetric kits: Serum Triglyceride Determination (Sigma Diagnostics, St. Louis, MO, USA) and NEFA-HR kit (Wako Chemicals GmbH, Neuss, Germany), according to manufacturer’s instructions, and myo-Inositol Assay Kit (Megazyme Ltd., Wicklow, Ireland), as previously adapted [27 (link)]. Glucose levels were measured in fresh blood by Accu-Check Glucometer (Roche Diagnostics, Barcelona, Spain).

Castillo P., Palou M., Yau-Qiu Z.X., Rodríguez A.M., Palou A, & Picó C. (2021). Myo-Inositol Supplementation in Suckling Rats Protects against Adverse Programming Outcomes on Hypothalamic Structure Caused by Mild Gestational Calorie Restriction, Partially Comparable to Leptin Effects. Nutrients, 13(9), 3257.

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Blood Glucose and Biomarker Analysis

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Blood samples were collected in EDTA tubes and centrifuged at 5000 rpm for 10 minutes at 4°C and kept at 220°C until further analysis. Blood glucose was monitored by the Accu-Check Avia monitoring system (Roche Diagnostics, Rotkreuz, Switzerland). Plasma levels of insulin (Mercodia Rat Insulin ELISA; Mercodia AB, Uppsala, Sweden) and acetaminophen (Acetaminophen Direct ELISA Kit; Immuneanalysis, Pomona, CA) were analyzed according to the manufacturer's instructions.

Larsen A.T., Sonne N., Andreassen K.V., Gehring K., Karsdal M.A, & Henriksen K. (2019). The Dual Amylin and Calcitonin Receptor Agonist KBP-088 Induces Weight Loss and Improves Insulin Sensitivity Superior to Chronic Amylin Therapy. The Journal of pharmacology and experimental therapeutics, 370(1).

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Insulin Assays and Tolerance Test

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Fasting and fed insulin: Insulin levels were measured in serum samples (in fed and 6-h fasting conditions, performed between 8:00 and 14:00) by using a rat insulin ELISA (Enzyme-Linked Immuno-Sorbent Assay) kit from Mercodia (Uppsala, Sweden).
Insulin tolerance test (ITT): In the last day of the last treatment week (23), rats were administered intraperitoneally with insulin (0.75 U/kg BW, Actrapid Novo Nordisk) following a 6-h fasting period. Blood glucose levels were obtained through a drop of blood from the tail vein before the bolus and 30, 60 and 120 min after, using the portable device Accu-Chek^® Aviva glucometer (Roche, Mannheim, Germany). The area under the curve (AUC) for the ITT was calculated by using the trapezoidal method, as previously described [24 (link)].

Nunes S., Alves A., Preguiça I., Barbosa A., Vieira P., Mendes F., Martins D., Viana S.D, & Reis F. (2020). Crescent-Like Lesions as an Early Signature of Nephropathy in a Rat Model of Prediabetes Induced by a Hypercaloric Diet. Nutrients, 12(4), 881.

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