Mouse insulin elisa

Manufactured by Mercodia

Sourced in Sweden, United States

The Mouse Insulin ELISA is a laboratory assay used to measure insulin levels in mouse samples. It is a quantitative enzyme-linked immunosorbent assay (ELISA) designed to determine the concentration of insulin in mouse serum, plasma, or other biological samples.

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Close spelling variants of this product

Mouse insulin ELISA kit (161 citations) Insulin (96 citations) Insulin ELISA (62 citations) Ultrasensitive Mouse Insulin ELISA (47 citations) Ultrasensitive Mouse Insulin ELISA kit (39 citations) ELISAs (9 citations) Mouse ultra sensitive insulin ELISA kit (4 citations) Mouse insulin enzyme-linked immunosorbent assay (ELISA) kit (3 citations) Ultrasensitive mouse insulin enzyme-linked immunosorbent assay (ELISA) kit (3 citations) Mouse and rat insulin ELISA (2 citations)

69 protocols using mouse insulin elisa

Metabolic effects of OP449 in mice

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Rag/MKR female mice, age 8–10 were allocated, 3–4 mice per cage, and treated intraperitoneally with 5mg/kg OP449 or vehicle, five times per week for three consecutive weeks. Body weight was measured weekly. Non-fasting glucose was measured in blood samples from the capillary tail vein once weekly, using a Bayer Contour Next Glucometer (Bayer, Mishawaka, IN). Glucose tolerance test was performed on mice fasted for 10 h. Glucose (1.5g/kg) was injected intraperitoneally. Capillary tail vein blood glucose levels were measured immediately before glucose injection at time 0 and at 15, 30, 60 and 120 min using Bayer Contour Next Glucometer (Bayer, Mishawaka, IN). Body composition was determined in non-anesthetized mice using the EchoMRI 3-in-1 NMR system (Echo Medical Systems, Houston, TX). At the end of the treatment period, fasting plasma and serum were collected for subsequent analysis.
Insulin levels were measured using the Mercodia Mouse Insulin ELISA (Mercodia AB, Upsala, Sweden). Serum triglyceride and cholesterol levels were determined using the Pointe Scientific Liquid Triglyceride and Liquid Cholesterol kits, respectively, per the manufacturer’s instructions (Pointe Scientific, Canton, MI).

Shlomai G., Zelenko Z., Antoniou I.M., Stasinopoulos M., Tobin-Hess A., Vitek M.P., LeRoith D, & Gallagher E.J. (2017). OP449 Inhibits Human Breast Cancer Growth without Adverse Metabolic Effects. Endocrine-related cancer, 24(10), 519-529.

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Glucose Tolerance Test in Mice

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For the GTT, d-glucose (2 g/kg) was injected intraperitoneally (i.p.) in mice anesthetized with midazolam (0.4 mg/mouse; Dormicum®; Hoffmann-La Roche) and a combination of fluanison (0.9 mg/mouse) and fentanyl (0.02 mg/mouse; Hypnorm®; Janssen, Beerse, Belgium). Mice were fasted 4 h before the GTT and kept on a heating pad for the entire time to maintain the body temperature. Blood (30 μl) was drawn retro-orbitally at 0, 15, 30, 60, and 120 min after glucose injection. Plasma was stored at − 80 °C until analysis. Glucose and insulin levels were determined by Amplex red glucose/glucose oxidase assay kit (Invitrogen™) and insulin ELISA (Mercodia Mouse Insulin ELISA; Mercodia AB, Uppsala, Sweden), respectively. Fasting blood collected from the tolerance test at time point 0 was used to obtain an insulin resistance index (Homeostatic Model Assessment of Insulin Resistance (HOMA-IR); HOMA-IR = (Glucose₀ in mmol/l) × (Insulin₀ in μg/l)/22.5) [30 (link)]. The cut-off value for insulin resistance was chosen based on the 90th percentile of the HOMA-IR values of the sham/CTRL diet group [31 (link)].

Elabi O.F., Cunha J.P., Gaceb A., Fex M, & Paul G. (2021). High-fat diet-induced diabetes leads to vascular alterations, pericyte reduction, and perivascular depletion of microglia in a 6-OHDA toxin model of Parkinson disease. Journal of Neuroinflammation, 18, 175.

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Glucose and Insulin Tolerance Evaluation

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We performed intraperitoneal glucose tolerance test (IPGTT) and intraperitoneal insulin tolerance test (IPITT) to evaluate glucose and insulin tolerance 2 weeks before the mice were sacrificed. Mice were injected intraperitoneally with 0.75 U/kg insulin after 4 h of fasting or 2.5 g/kg glucose after 12 h of fasting. Blood glucose from the tail tip was measured at 0, 30, 60, 90, and 120 min after injection using a Roche blood glucose monitoring system. Serum insulin levels were quantitatively measured by a commercial ELISA kit (Mercodia Mouse Insulin ELISA, 10-1247-10; Mercodia, Uppsala, Sweden). Insulin resistance index (HOMA-IR) was calculated using fasting serum glucose and fasting serum insulin according to the previously mentioned article (Yu et al., 2023 (link)).

Ye J., Chen J., Li Y., Sun L, & Lu H. (2023). Hepatocyte-specific knockout of HIF-2α cannot alleviate carbon tetrachloride-induced liver fibrosis in mice. PeerJ, 11, e15191.

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Multiparametric Serum Analysis in Mice

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Trunk serum was analysed by ELISA for levels of insulin (Mercodia Mouse Insulin ELISA; Cat No. 10-1247-01; Uppsala, Sweden), leptin (Crystal Chem Inc.; Cat. No. 90030; IL 60515, USA), C-peptide (Crystal Chem Inc.; Cat. No. 90050; IL 60515, USA), adiponectin (Crystal Chem Inc.; Cat. No. 80569; IL 60515, USA) and glucagon (Mercodia Glucagon ELISA – 10 μl; Cat No. 10-1281-01; Uppsala, Sweden). Trunk serum was analysed by enzymatic assay for glucose (Crystal Chem Inc.; Cat. No. 81692; IL 60515, USA) and glycated haemoglobin (HbA1c, Crystal Chem Inc.; Cat. No. 80310; IL 60515, USA) according to the protocols described by the manufacturers.

Ryan P.M., Patterson E., Kent R.M., Stack H., O’Connor P.M., Murphy K., Peterson V.L., Mandal R., Wishart D.S., Dinan T.G., Cryan J.F., Seeley R.J., Stanton C, & Ross R.P. (2017). Recombinant Incretin-Secreting Microbe Improves Metabolic Dysfunction in High-Fat Diet Fed Rodents. Scientific Reports, 7, 13523.

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Glucose Metabolism Monitoring in Mice

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Blood samples were obtained from the tail on Weeks 1, 4, 9, 12, and 16, and glucose was measured with a GlucoSure VIVO blood glucose meter (APEXBIO, Hsinchu, Taiwan). The area under the curve (AUC) was calculated from the glucose on Weeks 1, 4, 9, 12 and 16 using GraphPad Prism 6.0 (GraphPad software, San Diego, CA, USA). At the 16th week of the experimental period, plasma insulin levels were measured using Mercodia Mouse Insulin ELISA (Cat# 10-1247-01, Mercodia, Winston Salem, NC, USA), and the homeostasis model assessment (HOMA-IR) was calculated using the following equation: HOMA-IR = (glucose (mg/dL) × insulin (mIU/L))/405.

Tung Y.T., Zeng J.L., Ho S.T., Xu J.W., Lin I.H, & Wu J.H. (2021). Djulis Hull Improves Insulin Resistance and Modulates the Gut Microbiota in High-Fat Diet (HFD)-Induced Hyperglycaemia. Antioxidants, 11(1), 45.

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Plasma Biomarkers Quantification Protocol

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Blood samples were collected in EDTA tubes and centrifuged at 5,000 rpm for 10 min at 4°C. Plasma insulin levels (Mercodia mouse Insulin ELISA, Mercodia AB, Uppsala, Sweden) were analysed according to manufacturer instructions. ALT, AST, ALP, -TG, LDL, HDL, and chol levels were measured in-house (ADVIA® 1800, Siemens Healthineer, Germany). Basement membrane type IV collagen, PRO-C4 (Nordic Bioscience A/S, Herlev) was assessed by a validated competitive ELISA (Leeming et al., 2012) .A streptavidin-coated 96 well plate with the appropriate biotinylated peptide was incubated for 30 min in dark at 20°C while shaking (300 rpm) and subsequently washed in washing buffer. Twenty microliters of controls and samples was added to the wells together with 100µL horseradish peroxidase conjugated monoclonal antibody and incubated for 1 hour at 20°C in dark while shaking and washed in washing buffer. Hundred microliters of substrate solution were added to each well for 15 minutes left in the dark while shaking following 100µL of stopping solution. Plates were measured at 450nm with 650nm as reference.

Kayed A., Melander S.A., Khan S., Andreassen K.V., Karsdal M.A, & Henriksen K. (2023). The Effects of Dual GLP-1/Glucagon Receptor Agonists with Different Receptor Selectivity in Mouse Models of Obesity and Nonalcoholic Steatohepatitis. The Journal of pharmacology and experimental therapeutics, 384(3).

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Serum Biomarker Quantification in Mice

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Mouse Insulin ELISA (Cat # 10‐1247‐01, Mercodia, Uppsala, Sweden), mouse Adiponectin ELISA kit (Cat #: 80569, Crystal Chem, IL, USA), and mouse Leptin ELISA kit (Cat #: 90030, Crystal Chem) were used to determine insulin, adiponectin, and leptin amounts in the serum of WT and CD169‐DTR mice following the manufacturer’s instructions.

Chen Q., Lai S.M., Xu S., Tan Y., Leong K., Liu D., Tan J.C., Naik R.R., Barron A.M., Adav S.S., Chen J., Chong S.Z., Ng L.G, & Ruedl C. (2021). Resident macrophages restrain pathological adipose tissue remodeling and protect vascular integrity in obese mice. EMBO Reports, 22(8), e52835.

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Insulin Secretion Measurement in MIN6 Cells

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The MIN6 cells used were a gift of Prof. Miyazaki (Division of Stem Cell Regulation Research, Osaka University Graduate School of Medicine, Osaka Japan). Measurement of insulin secretion of MIN6 cells was performed according to the original report (20 (link)). Cells (ca. 5 × 10⁵/100 μl) were placed in 24- or 96-well plates (100–300 μl) and grown for 1 day in DMEM containing 25 mmol/l glucose and 10% FBS at 37°C with 5% CO₂. The cells were then washed twice in DMEM containing 0.5 mmol/l glucose and 10% FBS and cultured in this medium for 1 day. The medium was replaced with 0.5 ml of 0.5, 5.5, or 25 mmol/l glucose DMEM containing 5% FBS every 1 h.
All culture supernatants were collected after each incubation, centrifuged briefly to remove cell debris, and stored at −20°C before being studied using an enzyme-linked immunosorbent assay (mouse insulin ELISA; Mercodia, Uppsala, Sweden). The cells were harvested and a cell lysate was prepared with a mammalian protein extraction reagent (M-PER; Thermo Fisher Scientific, Waltham, MA, USA) for determination of cell protein.

Matsumura K., Tamasawa N, & Daimon M. (2018). Possible Insulinotropic Action of Apolipoprotein A–I Through the ABCA1/Cdc42/cAMP/PKA Pathway in MIN6 Cells. Frontiers in Endocrinology, 9, 645.

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Mouse Serum Insulin Quantification by ELISA

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Mouse serum was tested for insulin using the Mouse Insulin ELISA (Mercodia, cat# 10–1247-01) as described in the manufacturer’s instructions. Briefly, 10 µL of serum or calibrator was added to the wells, 100 µL of enzyme conjugate solution was added, and the plate was incubated for 2 h at room temperature at 700 rpm on a plate shaker. Each well was washed 6 times with 700 µL of wash buffer, and 200 µL of Substrate TMB was added to each well followed by a 15-min incubation at room temperature. Then, 50 µL of Stop Solution was added to each well, the plate was shaken for 5 s and then absorbance was measured at 450 nm using a SpectraMax i3x plate reader (Molecular Devices). Insulin concentrations were calculated based on calibrators provided in the kit.

Wilson M.R., Skalski H., Reske J.J., Wegener M., Adams M., Hostetter G., Hoffmann H.M., Bernard J.J., Bae-Jump V.L., Teixeira J.M, & Chandler R.L. (2022). Obesity alters the mouse endometrial transcriptome in a cell context-dependent manner. Reproductive Biology and Endocrinology : RB&E, 20, 163.

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Glucose Tolerance Assay in Mice

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IVGTTs were performed on 5 h fasted mice after 8 weeks of diet treatment. Blood samples were collected from anaesthetized mice (Hypnorm, 0.5 mg fluanisone, 0.02 mg fentanyl per mouse; Janssen, Beerse, Belgium; 0.25 mg midazolam per mouse; Dormicum, Hoffman LaRoche, Basel, Switzerland) from the retrobulbar, intraorbital, capillary plexus before d‐glucose injection (0.35 g/kg bw) to the tail vein. Additional blood samples were collected at 1, 5, 10, 20, 30, and 50 min from each mouse. Plasma was separated and stored at −20°C until it was analyzed for insulin with a mouse insulin ELISA (Mercodia, Uppsala, Sweden) and glucose (glucose oxidase method).

Omar B.A., Pacini G, & Ahrén B. (2014). Impact of glucose dosing regimens on modeling of glucose tolerance and β‐cell function by intravenous glucose tolerance test in diet‐induced obese mice. Physiological Reports, 2(5), e12011.

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