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Human, Mouse, & Rat Glucagon ELISA Kit - Quantikine

Catalog #: DGCG0
15 Citations 1 Review Datasheet / COA / SDS
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QC100: Quantikine Immunoassay Control Set 869 for Glucagon
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Multi-species Glucagon ELISA Standard Curve
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Citations (15)
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Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Glucagon Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
14.7 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural Glucagon.  This assay cross-reacts <12% with Oxyntomodulin.
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Glucagon Immunoassay is a 4.5 hour solid-phase ELISA designed to measure Glucagon in cell culture supernates, serum, and plasma. It contains natural porcine Glucagon as the standard. The antibodies were raised against a human Glucagon synthetic peptide. This immunoassay has been shown to accurately quantitate human, mouse, rat, and porcine Glucagon.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 315 618 1024 354 653 1080
Standard Deviation 10.3 22 27.6 30.7 37.8 63.6
CV% 3.3 3.6 2.7 8.7 5.8 5.9

Recovery

The recovery of Glucagon spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 98 91-108
Human EDTA Plasma (n=4) 98 86-107
Human Heparin Plasma (n=4) 96 88-100
Human Serum (n=4) 102 95-111
Mouse Serum (n=2) 99 92-106
Porcine Serum (n=2) 97 91-107
Rat Serum (n=1) 93 86-101

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of Glucagon were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Multi-species Glucagon ELISA Linearity
Multi-species Glucagon ELISA Linearity

Scientific Data

N/A Multi-species Glucagon ELISA Standard Curve View Larger

Multi-species Glucagon ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Glucagon

Glucagon is a 29 amino acid (aa) peptide produced by the pancreas that plays a critical role in glucose metabolism and homeostasis (1-4). The Glucagon precursor mRNA is expressed by alpha cells ( alpha -cells) of the pancreas, L cells of the intestine, and in the brain (1, 2). Only the pancreatic alpha -cells express the prohormone convertase PC2, also called PCSK2, which is required to produce Glucagon (2). Intestinal L cells instead express the prohormone convertase PC1, which processes the precursor to the Glucagon-overlapping peptides glicentin and oxyntomodulin. L cells also produce two Glucagon-like peptides, GLP-1 and GLP-2 that are derived from the same Glucagon precursor and influence glucose metabolism, but do not share any common sequence with Glucagon (1, 2). The aa sequence of the mature Glucagon peptide is identical in human, mouse, rat, pig, dog, horse, cow, sheep, and Xenopus. 

In normal metabolism, Glucagon is secreted in response to low blood glucose (hypoglycemia) and downregulated in response to high blood glucose (hyperglycemia). Although Glucagon binding sites are found in liver, brain, pancreas, kidney, intestine, and adipose tissue, the main activity of Glucagon receptors occurs in the liver, where Glucagon stimulates gluconeogenesis and glycogenolysis, thereby increasing blood glucose (1-4). It is particularly important that the brain receive sufficient glucose, since it is unable to store more than a minute quantity. Therefore the release of Glucagon from alpha -cells is under control by both hormones and neurotransmitters, and is very responsive to circulating glucose concentration. Insulin, and/or the zinc that islet beta cells secrete with it, downregulates Glucagon secretion in intact islets (5, 6). Glucagon secretion is also downregulated by the neurotransmitter gamma -aminobutyric acid (GABA), somatostatin produced by islet δ-cells, and GLP-1, but is enhanced by the neurotransmitter L-glutamate, amino acids (especially arginine), and Glucagon itself (2-4, 7). Through receptors on the alpha -cells, these substances affect potassium, sodium, and calcium channel activity and alter intracellular calcium concentration (2-4). Glucose suppression of Glucagon secretion is probably indirect, acting through paracrine signals from other islet cells (8). 
Like insulin, Glucagon is dysregulated in type 2 diabetes (T2D) and contributes to its pathology (2-4). Glucagon secretion is less responsive to insulin-mediated suppression in times of high circulating glucose, causing glucagonemia, and increasing the risk of hyperglycemia. Glucagon is also regulated by some of the same messengers that regulate insulin (10-12). Leptin inhibits alpha -cell glucagon secretion and stimulates beta -cell insulin secretion, but glucagon blunts the leptin-mediated insulin secretion (10). Islet alpha -cells express ghrelin receptors and respond to ghrelin by increasing Glucagon secretion (11). Glucocorticoids, activated by 11 beta -HSD1, depress Glucagon secretion in hypoglycemia and insulin secretion in hyperglycemia (12). Although genetic polymorphisms of the Glucagon receptor are associated with T2D, downregulation of Glucagon secretion or deletion of the Glucagon receptor in mice that are susceptible to T2D actually improves glycemic control (13, 14).

Entrez Gene IDs:
2641 (Human); 14526 (Mouse); 24952 (Rat)
Alternate Names:
GCG; glicentin-related polypeptide; GLP1; GLP-1; Glucagon; glucagon-like peptide 1; glucagon-like peptide 2; GRPP
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

The conjugate must be kept cold during use. Bring all other reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
  3.   Wash and aspirate the plate a total of 2 times with Wash Buffer prior to assay.
    4. 150 µL Assay Diluent
  4.   Add 150 µL of Assay Diluent to each well.
    5. 50 µL Standard, Control, or Sample
  5.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
  6.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    7. 200 µL cold Conjugate
  7.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
  8.   Aspirate and wash 4 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Glucagon Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

15 Citations: Showing 1 - 10
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  1. Gut AstA mediates sleep deprivation-induced energy wasting in Drosophila
    Authors: Li, Y;Zhou, X;Cheng, C;Ding, G;Zhao, P;Tan, K;Chen, L;Perrimon, N;Veenstra, JA;Zhang, L;Song, W;
    Cell discovery
    Species: Mouse
    Sample Types: Cell Culture Supernates, Serum
  2. Low glycaemic diets alter lipid metabolism to influence tumour growth
    Authors: EC Lien, AM Westermark, Y Zhang, C Yuan, Z Li, AN Lau, KM Sapp, BM Wolpin, MG Vander Hei
    Nature, 2021-10-20;0(0):.
    Species: Human
    Sample Types: Plasma
  3. Intervention of Gastrodin in Type 2 Diabetes Mellitus and Its Mechanism
    Authors: Y Bai, K Mo, G Wang, W Chen, W Zhang, Y Guo, Z Sun
    Frontiers in Pharmacology, 2021-09-16;12(0):710722.
    Species: Rat
    Sample Types: Serum
  4. Glucagon receptor antagonism promotes the production of gut proglucagon-derived peptides in diabetic mice
    Authors: S Lang, R Wei, T Wei, L Gu, J Feng, H Yan, J Yang, T Hong
    Peptides, 2020-06-16;131(0):170349.
    Species: Mouse
    Sample Types: Plasma
  5. Low-Energy Extracorporeal Shock Wave Ameliorates Streptozotocin Induced Diabetes and Promotes Pancreatic Beta Cells Regeneration in a Rat Model
    Authors: CC Hsiao, CC Lin, YS Hou, JY Ko, CJ Wang
    Int J Mol Sci, 2019-10-05;20(19):.
    Species: Rat
    Sample Types: Tissue Lysates
  6. Sodium butyrate reduces high-fat diet-induced non-alcoholic steatohepatitis through upregulation of hepatic GLP-1R expression
    Authors: D Zhou, YW Chen, ZH Zhao, RX Yang, FZ Xin, XL Liu, Q Pan, H Zhou, JG Fan
    Exp. Mol. Med., 2018-12-03;50(12):157.
    Species: Human
    Sample Types: Serum
  7. Alpha cell function interacts with diet to modulate prediabetes and Type 2 diabetes
    Authors: I Roncero-Ra, R Jimenez-Lu, JF Alcala-Dia, C Vals-Delga, AP Arenas-Lar, OA Rangel-Zuñ, A Leon-Acuña, MM Malagon, J Delgado-Li, P Perez-Mart, JM Ordovas, A Camargo, J Lopez-Mira
    J. Nutr. Biochem., 2018-09-01;62(0):247-256.
    Species: Human
    Sample Types: Plasma
  8. Metabolic regulation of GLP-1 and PC1/3 in pancreatic ?-cell line
    Authors: V Sancho, G Daniele, D Lucchesi, R Lupi, A Ciccarone, G Penno, C Bianchi, A Dardano, R Miccoli, S Del Prato
    PLoS ONE, 2017-11-09;12(11):e0187836.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  9. Involvement of stanniocalcins in the deregulation of glycaemia in obese mice and type 2 diabetic patients
    Authors: JJ López, I Jardín, CC Chamorro, ML Duran, MJ Tarancón R, M Reyes Pana, F Jiménez, R Montero, MJ González, M Martínez, MJ Hernández, JM Brull, AJ Corbacho, E Delgado, MP Granados, L Gómez-Gord, JA Rosado, PC Redondo
    J. Cell. Mol. Med., 2017-10-09;0(0):.
    Species: Mouse
    Sample Types: Plasma
  10. Hypothalamic Ventromedial Lin28a Enhances Glucose Metabolism in Diet-Induced Obesity
    Authors: JD Kim, C Toda, CM Ramírez, C Fernández-, S Diano
    Diabetes, 2017-05-26;0(0):.
    Species: Mouse
    Sample Types: Plasma
  11. Effects of Three Commercially Available Sports Drinks on Substrate Metabolism and Subsequent Endurance Performance in a Postprandial State
    Authors: L Qin, QR Wang, ZL Fang, T Wang, AQ Yu, YJ Zhou, Y Zheng, MQ Yi
    Nutrients, 2017-04-12;9(4):.
    Species: Human
    Sample Types: Serum
  12. Inhibition of the glucose transporter SGLT2 with dapagliflozin in pancreatic alpha cells triggers glucagon secretion.
    Authors: Bonner C, Kerr-Conte J, Gmyr V, Queniat G, Moerman E, Thevenet J, Beaucamps C, Delalleau N, Popescu I, Malaisse W, Sener A, Deprez B, Abderrahmani A, Staels B, Pattou F
    Nat Med, 2015-04-20;21(5):512-7.
    Species: Human, Mouse
    Sample Types: Cell Culture Supernates
  13. Vitamin A deficiency causes hyperglycemia and loss of pancreatic beta-cell mass.
    Authors: Trasino S, Benoit Y, Gudas L
    J Biol Chem, 2014-12-01;290(3):1456-73.
    Species: Mouse
    Sample Types: Serum
  14. Hypothalamic prolyl endopeptidase (PREP) regulates pancreatic insulin and glucagon secretion in mice.
    Authors: Kim J, Toda C, D'Agostino G, Zeiss C, DiLeone R, Elsworth J, Kibbey R, Chan O, Harvey B, Richie C, Savolainen M, Myohanen T, Jeong J, Diano S
    Proc Natl Acad Sci U S A, 2014-07-28;111(32):11876-81.
    Species: Mouse
    Sample Types: Tissue Homogenates
  15. A novel oral dual amylin and calcitonin receptor agonist (KBP-042) exerts antiobesity and antidiabetic effects in rats.
    Authors: Andreassen K, Feigh M, Hjuler S, Gydesen S, Henriksen J, Beck-Nielsen H, Christiansen C, Karsdal M, Henriksen K
    Am J Physiol Endocrinol Metab, 2014-05-06;307(1):E24-33.
    Species: Rat
    Sample Types: Plasma

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Glucagon Quantikine ELISA Kit
By Anonymous on 07/19/2017
Sample Tested: Serum

works for mouse serum