• 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 (25 uL), EDTA Plasma (25 uL), Heparin Plasma (25 uL)
  • Sensitivity
    13.4 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant mouse and rat FGF-21
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse/Rat FGF-21 immunoassay is a 4.5 hour solid-phase ELISA designed to measure FGF-21 in mouse or rat cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse FGF-21 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse FGF-21. Results obtained using natural mouse or rat FGF-21 showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural FGF-21.

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
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean84.422675494.5231745
Standard Deviation5.249.619.76.6110.831
CV%6.24.22.674.74.2

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean1183171032120302977
Standard Deviation7.3513.525.28.911865.9
CV%6.24.32.47.466.7

Recovery

The recovery of FGF-21 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Samples (n=6) 96 88-101
Mouse EDTA Plasma (n=4) 97 86-104
Mouse Heparin Plasma (n=4) 93 83-101
Mouse Serum (n=4) 95 83-103
Rat Cell Culture Samples (n=2) 91 87-95
Rat EDTA Plasma (n=4) 101 93-119
Rat Heparin Plasma (n=4) 97 94-104
Rat Serum (n=4) 100 93-108
Linearity
To assess the linearity of the assay, samples spiked with high concentrations of FGF-21 in each matrix were diluted with the appropriate Calibrator Diluent and assayed.
Mouse/Rat FGF-21 Quantikine ELISA Kit
Mouse/Rat FGF-21 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: FGF-21
FGF-21 (Fibroblast Growth Factor 21) is a member of an FGF subfamily that also includes FGF-19 and FGF-23. FGF-19 subfamily members lack a heparin binding domain, enabling them to freely circulate as endocrine factors and diffuse within tissues. FGF-21 signals through Klotho beta in complex with FGF receptors. It regulates cellular metabolism including glucose uptake in adipocytes and cellular sensitivity to insulin. It is basally expressed in the pancreas, thymus, liver, and adipose tissue and is upregulated in liver, muscle, and fat in obesity and diabetes. FGF-21 also plays a role in cell survival and proliferation, mesenchymal stem cell differentiation, circadian rhythm, and controlling reproductive capacity.
    • Long Name
      Fibroblast Growth Factor 21
    • Entrez Gene IDs
      26291 (Human); 56636 (Mouse);
    • Alternate Names
      FGF21; FGF-21; fibroblast growth factor 21;
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    Bring all 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. 50 µL Assay Diluent
    4.   Add 50 µL of Assay Diluent to each well.

    5. 50 µL Standard, Control, or Sample
    6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

    8. 100 µL Conjugate
    9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    10.   Aspirate and wash 5 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

    13. 100 µL Stop Solution
    14.   Add 100 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.

    Showing Results 1 - 10 of 13
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    Species
    Sample Type
    1. Hyperlipidemia and hepatitis in liver-specific CREB3L3 knockout mice generated using a one-step CRISPR/Cas9 system
      Authors: Yoshimi Nakagawa
      Sci Rep, 2016;6(0):27857.
      Species: Mouse
      Sample Type: Plasma
    2. Single ingestion of soy ?-conglycinin induces increased postprandial circulating FGF21 levels exerting beneficial health effects
      Authors: Tsutomu Hashidume
      Sci Rep, 2016;6(0):28183.
      Species: Mouse
      Sample Type: Serum
    3. CREBH-FGF21 axis improves hepatic steatosis by suppressing adipose tissue lipolysis
      Authors: Jong-Gil Park
      Sci Rep, 2016;6(0):27938.
      Species: Mouse
      Sample Type: Plasma
    4. The Orphan Nuclear Receptor ERR? Regulates Hepatic CB1 Receptor-Mediated Fibroblast Growth Factor 21 Gene Expression
      PLoS ONE, 2016;11(7):e0159425.
      Species: Mouse
      Sample Type: Serum
    5. Intramyocardial Adipose-Derived Stem Cell Transplantation Increases Pericardial Fat with Recovery of Myocardial Function after Acute Myocardial Infarction
      Authors: Jong-Ho Kim
      PLoS ONE, 2016;11(6):e0158067.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    6. A muscle-liver-fat signalling axis is essential for central control of adaptive adipose remodelling.
      Authors: Shimizu N, Maruyama T, Yoshikawa N, Matsumiya R, Ma Y, Ito N, Tasaka Y, Kuribara-Souta A, Miyata K, Oike Y, Berger S, Schutz G, Takeda S, Tanaka H
      Nat Commun, 2015;6(0):6693.
      Species: Mouse
      Sample Type: Plasma
    7. Hepatic mitogen-activated protein kinase phosphatase 1 selectively regulates glucose metabolism and energy homeostasis.
      Authors: Lawan A, Zhang L, Gatzke F, Min K, Jurczak M, Al-Mutairi M, Richter P, Camporez J, Couvillon A, Pesta D, Roth Flach R, Shulman G, Bennett A
      Mol Cell Biol, 2015;35(1):26-40.
      Species: Mouse
      Sample Type: Serum
    8. Skeletal muscle mitochondrial uncoupling drives endocrine cross-talk through the induction of FGF21 as a myokine.
      Authors: Keipert S, Ost M, Johann K, Imber F, Jastroch M, van Schothorst E, Keijer J, Klaus S
      Am J Physiol Endocrinol Metab, 2014;306(5):E469-82.
      Species: Mouse
      Sample Type: Plasma
    9. Long-term low carbohydrate diet leads to deleterious metabolic manifestations in diabetic mice.
      Authors: Handa K, Inukai K, Onuma H, Kudo A, Nakagawa F, Tsugawa K, Kitahara A, Moriya R, Takahashi K, Sumitani Y, Hosaka T, Kawakami H, Oyadomari S, Ishida H
      PLoS ONE, 2014;9(8):e104948.
      Species: Mouse
      Sample Type: Serum
    10. Somatic progenitor cell vulnerability to mitochondrial DNA mutagenesis underlies progeroid phenotypes in Polg mutator mice.
      Authors: Ahlqvist KJ, Hamalainen RH, Yatsuga S, Uutela M, Terzioglu M, Gotz A, Forsstrom S, Salven P, Angers-Loustau A, Kopra OH, Tyynismaa H, Larsson NG, Wartiovaara K, Prolla T, Trifunovic A, Suomalainen A
      Cell Metab., 2012;15(1):100-9.
      Species: Mouse
      Sample Type: Serum
    11. Activation of the farnesoid x receptor induces hepatic expression and secretion of fibroblast growth factor 21.
      Authors: Cyphert HA, Ge X, Kohan AB, Salati LM, Zhang Y, Hillgartner FB
      J. Biol. Chem., 2012;287(30):25123-38.
      Species: Rat
      Sample Type: Plasma
    12. Fasting-induced FGF21 is repressed by LXR activation via recruitment of an HDAC3 corepressor complex in mice.
      Authors: Archer A, Venteclef N, Mode A, Pedrelli M, Gabbi C, Clement K, Parini P, Gustafsson J, Korach-Andre M
      Mol Endocrinol, 2012;26(12):1980-90.
      Species: Mouse
      Sample Type: Serum
    13. Paradoxical regulation of human FGF21 by both fasting and feeding signals: is FGF21 a nutritional adaptation factor?
      Authors: Uebanso T, Taketani Y, Yamamoto H, Amo K, Ominami H, Arai H, Takei Y, Masuda M, Tanimura A, Harada N, Yamanaka-Okumura H, Takeda E
      PLoS ONE, 2011;6(8):e22976.
      Species: Human
      Sample Type: Cell Culture Supernates
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