• 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.
Control Available
QC100, Quantikine Immunoassay Control Set 869 for Glucagon - Please Inquire
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.

Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean31561810243546531080
Standard Deviation10.32227.630.737.863.6
CV%3.33.62.78.75.85.9

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean31561810243546531080
Standard Deviation10.32227.630.737.863.6
CV%3.33.62.78.75.85.9

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.
Glucagon Quantikine ELISA Kit
Glucagon Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Glucagon
Glucagon is a pancreatic peptide hormone that plays a critical role in glucose metabolism and homeostasis. Mature Glucagon is generated from its precursor by the prohormone convertase PC2/PCSK2 in pancreatic alpha-cells. Glucagon is secreted in response to low blood glucose (hypoglycemia) and downregulated in response to high blood glucose (hyperglycemia). Glucagon production is under the control of both hormones and neurotransmitters. Glucagon inhibits Insulin secretion, and vice versa. Glucagon stimulates hepatic gluconeogenesis and glycogenolysis, thereby increasing blood glucose. Like insulin, Glucagon is dysregulated in type 2 diabetes (T2D) and contributes to its pathology.
    • Entrez Gene IDs
      2641 (Human); 14526 (Mouse); 24952 (Rat);
    • Alternate Names
      GCG; glicentin-related polypeptide; Glucagon; GRPP;
    Related Research Areas
    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
    5.   Add 150 µL of Assay Diluent to each well.

    6. 50 µL Standard, Control, or Sample
    7.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
    8.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    9. 200 µL cold Conjugate
    10.   Add 200 µL of cold Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 1 hour.
    11.   Aspirate and wash 4 times.

    12. 200 µL Substrate Solution
    13.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    14. 50 µL Stop Solution
    15. 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:

    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.

    4 Citations: Showing 1 - 4
    Filter your results:

    Species
    Sample Type
    1. Vitamin A deficiency causes hyperglycemia and loss of pancreatic beta-cell mass.
      Authors: Trasino S, Benoit Y, Gudas L
      J Biol Chem, 2015;290(3):1456-73.
      Species: Mouse
      Sample Type: Serum
    2. 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;21(5):512-7.
      Species: Human
      Sample Type: Cell Culture Supernates
    3. 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;307(1):E24-33.
      Species: Rat
      Sample Type: Plasma
    4. 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;111(32):11876-81.
      Species: Mouse
      Sample Type: Tissue Homogenates
    ELISA Controls
    Description Application Cat# Citations Images  

    Quantikine Immunoassay Control Set 869 for Glucagon

    Ctrl QC100
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    Supplemental ELISA Products
    Description Application Cat# Citations Images  

    Quantikine ELISA Wash Buffer 1

    ELISA WA126 6
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    Average Rating: 5 (Based on 1 review)

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