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Glucagon quantikine elisa kit

Manufactured by R&D Systems
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Sourced in France, United States

The Glucagon Quantikine ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of glucagon levels in biological samples. The kit utilizes a monoclonal antibody specific for glucagon that has been pre-coated onto a microplate. Samples and standards are pipetted into the wells, and any glucagon present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody specific for glucagon is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of glucagon bound. The intensity of the color is measured by a spectrophotometer.

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Lab products found in correlation

Most reagents, by Merck Group (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Cm h2dcfda, by Thermo Fisher Scientific (1 mentions) Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Rat mouse insulin elisa kit, by Merck Group (1 mentions) Chemix 180, by Sysmex (1 mentions) Ultra sensitive murine insulin elisa, by Mercodia (1 mentions) Glp 1 eia kit, by Merck Group (1 mentions) Bradford assay dc protein assay, by Bio-Rad (1 mentions) Cholesterol fluorometric assay kit, by Cayman Chemical (1 mentions) Lactate colorimetric fluorometric assay kit, by Abcam (1 mentions) Atp assay kit, by Abcam (1 mentions) Glucose colorimetric assay kit, by Cayman Chemical (1 mentions) Acetyl coenzyme a assay kit, by Merck Group (1 mentions) Mouse insulin elisa kit, by Thermo Fisher Scientific (1 mentions) Alt gpt reagent, by Thermo Fisher Scientific (1 mentions) Ast got reagent, by Thermo Fisher Scientific (1 mentions)

8 protocols using glucagon quantikine elisa kit

1

Immunoblotting and Gene Expression Analysis

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Most reagents were purchased from Sigma-Aldrich (St. Louise, MO). PC1/3, PC2 or Actin first antobodies and all HRP-conjugated secondary antibodies were purchased from Santa Cruz Biotechnologies (Dallas, TX). Pre-developed TaqMan gene expression assays, TaqMan Universal PCR Master Mix, CM-H2DCFDA, TaqMan Universal PCR Master Mix and Trizol were from Life Technologies (Grand Island, NY). iScript cDNA Synthesis kit, 4–20% Tris-Glycine gels precast gels and Trans-blot Tranfer Nitrocellulose membrane 0.20 μm were purchased from BioRad Laboratories Inc. (Hercules, CA). GLP-1 total and active form ELISA Kit were from Merck Millipore (Darmstadt, Germany). Glucagon quantikine ELISA Kit was from R&D Systems (Adbingdon, UK).
Sancho V., Daniele G., Lucchesi D., Lupi R., Ciccarone A., Penno G., Bianchi C., Dardano A., Miccoli R, & Del Prato S. (2017). Metabolic regulation of GLP-1 and PC1/3 in pancreatic α-cell line. PLoS ONE, 12(11), e0187836.
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2

Metabolic Biomarkers in Serum Analysis

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Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and fasting blood glucose were measured using an automated analyzer (Sysmex CHEMIX-180, Japan). Insulin (Rat/Mouse Insulin ELISA Kit, Merck-Millipore) and GLP-1 (Glucagon Quantikine ELISA Kit, R&D Systems) in the serum were measured using an enzyme-linked immunosorbent assay. A homeostatic model assessment of insulin resistance (HOMA-IR) and the insulin sensitive index (ISI) were calculated.
Zhou D., Chen Y.W., Zhao Z.H., Yang R.X., Xin F.Z., Liu X.L., Pan Q., Zhou H, & Fan J.G. (2018). Sodium butyrate reduces high-fat diet-induced non-alcoholic steatohepatitis through upregulation of hepatic GLP-1R expression. Experimental & Molecular Medicine, 50(12), 157.
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3

Measuring Islet Hormones by ELISA

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Commercially available ELISAs were used to measure insulin (Ultra-sensitive murine insulin ELISA, Mercodia), glucagon (Glucagon Quantikine ELISA Kit, R&D Systems) and GLP-1 (GLP-1 EIA Kit, Sigma-Aldrich) concentrations according to the manufacturers’ instructions. Further details on the processing of plasma and pancreatic protein extract for assessment of islet hormone concentrations is given in the methods section of the ESM.
Khan D., Kelsey R., Maheshwari R.R., Stone V.M., Hasib A., Manderson Koivula F.N., Watson A., Harkin S., Irwin N., Shaw J.A., McClenaghan N.H., Venglovecz V., Ébert A., Flodström-Tullberg M., White M.G, & Kelly C. (2019). Short-term CFTR inhibition reduces islet area in C57BL/6 mice. Scientific Reports, 9, 11244.
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4

Glucagon Secretion in Pancreatic α-Cells

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Pancreatic α-TC1 clone 6 cell line (ATCC, Middlesex, UK) were grown in DMEM with L-glutamine (modified to contain 16.7 mmol/L glucose and 1.5 g/L sodium bicarbonate) supplemented with 10% (vol/vol) inactivated foetal calf serum and 1% (vol/vol) antibiotics-antimycotics, 15 mmol/L HEPES, 0.1 mmol/L nonessential amino acids, and 0.02% BSA, and maintained in a humidified atmosphere of 5% CO2 in air at 37°C. Cells were subcultured once a week and the medium was replaced at least twice weekly. α-TC1 cells were treated with IGF-1 (5, 10, 50 or 100nM) for 24 h, Akt inhibitor VIII (210nM) for 2 h and/or phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 (40μM) for 30 min, as indicated. For glucagon secretion determinations, supernatants were collected from α-TC1 cells and glucagon secretion was measured using the Glucagon Quantikine ELISA Kit (R&D Systems Europe, Lille, France) according to the manufacturer's instructions. For normalization purposes, total protein amount was determined with the Bradford assay (DC Protein Assay; Bio-Rad, Hercules, CA, USA) according to the manufacturer's instructions.
Mancuso E., Mannino G.C., Fatta C.D., Fuoco A., Spiga R., Andreozzi F, & Sesti G. (2017). Insulin-like growth factor-1 is a negative modulator of glucagon secretion. Oncotarget, 8(31), 51719-51732.
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5

Metabolic Biomarker Analysis in Liver

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Liver TG, serum TG and serum cholesterol were measured with Triglyceride Colorimetric Assay Kit (Cayman Chemical #10010303) and Cholesterol Fluorometric Assay Kit (Cayman Chemical #10007640), respectively, according to manufacturer protocols. Serum insulin and glucagon concentrations were determined by Mouse Insulin ELISA KIT (Invitrogen, EMMINS) and Glucagon Quantikine ELISA Kit (R&D Systems, DGCG0), respectively, following manufacturer protocols. Liver NADPH and NADP were measured by NADP/NADPH-Glo™ Assays (Promega #G9081) according to manufacturer’s protocol. ALT and AST assays were performed with ALT(GPT) Reagent (Thermo Scientific™, TR71121) and AST/GOT Reagent (Thermo Scientific™, TR70121), respectively, according to the manufacturer’s protocol. ATP, Acetyl-CoA and G6P concentrations were determined by ATP assay kit (Abcam, ab83355), Acetyl-Coenzyme A Assay Kit (Sigma, MAK039) and PicoProbe™ Glucose-6-Phosphate Fluorometric Assay Kit (Biovision, K687), respectively according to the manufacturers protocols. Glucose and lactate were measured with Glucose colorimetric assay kit (Cayman Chemical #10009582) and Lactate Colorimetric/Fluorometric Assay Kit (Biovision, K607), respectively, according to the manufacturer’s protocols.
Gu L., Zhu Y., Watari K., Lee M., Liu J., Perez S., Thai M., Mayfield J.E., Zhang B., Rocha K.C., Li F., Kim L.C., Jones A.C., Wierzbicki I.H., Liu X., Newton A.C., Kisseleva T., Lee J.H., Ying W., Gonzalez D.J., Saltiel A.R., Simon M.C, & Karin M. (2023). Fructose-1,6-bisphosphatase is a nonenzymatic safety valve that curtails AKT activation to prevent insulin hyperresponsiveness. Cell metabolism, 35(6), 1009-1021.e9.
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6

Serum Biomarker Analysis in Mice

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At the end of 8 weeks, all mice were sacrificed by cervical dislocation. Serum was collected from the whole blood by centrifugation (12,000 rpm, 4°C, for 20 min). Serum insulin, glucagon, and leptin were respectively measured with an Ultra Sensitive Mouse Insulin ELISA kit (Crystal Chem, Downers Grove, IL, USA), Glucagon Quantikine ELISA kit (R&D Systems, Minneapolis, MN, USA), and Mouse/Rat Leptin Quantikine ELISA kit (R&D Systems) according to the manufacturer's protocols. The levels of serum glucose, triglyceride, total cholesterol, low-density lipoprotein (LDL)/very low-density lipoprotein (VLDL) cholesterol and high-density lipoprotein (HDL) cholesterol were measured using quantification kits from BioVision (Milpitas, CA, USA).
Kim O.K., Nam D.E., Jun W, & Lee J. (2015). Cudrania tricuspidata water extract improved obesity-induced hepatic insulin resistance in db/db mice by suppressing ER stress and inflammation. Food & Nutrition Research, 59, 10.3402/fnr.v59.29165.
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7

Insulin and Glucagon Quantification

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Serum concentrations of insulin or glucagon were determined using the Insulin ELISA kit (Mercodia Mouse Insulin ELISA #10-1247-10), and the Glucagon Quantikine ELISA kit (R&D Systems #DGCG0), respectively, according to the manufacturer’s instructions. HOMA I-R was calculated as glucose (mMol/L) X Insulin (mIU/L) /22.5.
Senatus L., Egaña-Gorroño L., López-Díez R., Bergaya S., Aranda J.F., Amengual J., Arivazhagan L., Manigrasso M.B., Yepuri G., Nimma R., Mangar K.N., Bernadin R., Zhou B., Gugger P.F., Li H., Friedman R.A., Theise N.D., Shekhtman A., Fisher E.A., Ramasamy R, & Schmidt A.M. (2023). DIAPH1 mediates progression of atherosclerosis and regulates hepatic lipid metabolism in mice. Communications Biology, 6, 280.
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8

Analytical Methods for Metabolic Biomarkers

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Before laboratory analysis, retrospectively identified frozen plasma samples were thawed at room temperature, centrifuged for 2 min at 4000 rpm, and then treated as per the individualized assay kit instructions. The Randox Daytona Plus RX series analyzer (Randox Laboratories Ltd.®, Crumlin, UK) was used for in vitro determination of β-hydroxybutyrate ([β-OHB] RB4067). C-peptide was quantified using Invitron's Molecular Light Technology™ chemiluminescence assay (IV2-004; Invitron Ltd.®, Monmouth, UK). Proinsulin was quantified using Invitron's Molecular Light Technology™ chemiluminescence intact assay (IV2-002, Invitron Ltd.). Glucagon was quantified using the R&D Systems® (R&D Systems, Inc., Minneapolis, MN) Glucagon Quantikine ELISA Kit (product no. DGCG0). The Eagle Biosciences (Eagle Biosciences, Inc., Amherst, NH) BI-CAT Epinephrine & Norepinephrine ELISA Assay Kit (product no. ECT31-K02) was used for the determination of catecholamines. Trial-day venous BG and blood lactate (BLa) concentrations were measured via the fully enzymatic-amperometric method (Biosen C-Line; EKF-diagnostic GmbH, Barleben, Germany).
McCarthy O., Pitt J., Eckstein M.L., Moser O., Bain S.C, & Bracken R.M. (2021). Pancreatic β-Cell Function Is Associated with Augmented Counterregulation to In-Exercise Hypoglycemia in Type 1 Diabetes. Medicine and science in sports and exercise, 53(7).
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