Mouse/Rat GDF-15 Quantikine ELISA Kit

(5 citations)
(1 Review)
  
  • 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), Urine (10 uL)
  • Sensitivity
    2.2 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural mouse and rat GDF-15, recombinant mouse mature GDF-15, precursor GDF-15, and GDF-15/BMP-2 heterodimer. It does not detect monomeric recombinant mouse GDF-15.
  • 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 GDF-15 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse and rat GDF-15 in cell culture supernates, serum, plasma, and urine. It contains E. coli-expressed recombinant mouse GDF-15 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse or rat GDF-15 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse and rat GDF-15.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean25.66019026.763.1196
Standard Deviation0.691.425.491.622.8410.7
CV%2.72.42.96.14.55.5

Recovery

The recovery of GDF-15 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Media (n=4) 102 91-110
Mouse EDTA Plasma (n=4) 95 81-109
Mouse Heparin Plasma (n=4) 95 87-104
Mouse Serum (n=4) 91 82-101
Mouse Urine (n=4) 97 88-106
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of GDF-15 in each matrix were diluted with Calibrator Diluent and assayed.
Mouse/Rat GDF-15 Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: GDF-15
Growth/differentiation factors (GDF-1 to GDF-15) are members of the BMP family of TGF-beta superfamily proteins. They are produced as inactive preproproteins which are then cleaved and assembled into active secreted homodimers. GDF dimers are disulfide-linked with the exception of GDF-3 and -9. GDF proteins are important during embryonic development, particularly in the skeletal, nervous, and muscular systems.
    • Long Name
      Growth Differentiation Factor 15
    • Entrez Gene IDs
      9518 (Human); 23886 (Mouse); 29455 (Rat);
    • Alternate Names
      GDF15; GDF-15; growth differentiation factor 15; growth/differentiation factor 15; Macrophage inhibitory cytokine 1; MIC-1; MIC-1NSAID-activated gene 1 protein; MIC1Prostate differentiation factor; NAG-1; NAG-1NSAID-regulated gene 1 protein; NSAID (nonsteroidal inflammatory drug)-activated protein 1; PDF; PDFGDF-15; PLAB; PLABNRG-1; Placental bone morphogenetic protein; Placental TGF-beta; PTGF-beta; PTGFBPTGF-beta;
    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 on a horizontal orbital microplate shaker.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 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 on the shaker.
    10.   Aspirate and wash 4 times.

    11. 100 µL Substrate Solution
    12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. 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.

    5 Citations: Showing 1 - 5
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    Species
    Sample Type
    1. Oral Treatment with the Ghrelin Receptor Agonist HM01 Attenuates Cachexia in Mice Bearing Colon-26 (C26) Tumors
      Authors: FO Villars, C Pietra, C Giuliano, TA Lutz, T Riediger
      Int J Mol Sci, 2017;18(5):.
      Species: Mouse
      Sample Type: Plasma
    2. GDF15 is elevated in mice following retinal ganglion cell death and in glaucoma patients
      Authors: N Ban, CJ Siegfried, JB Lin, YB Shui, J Sein, W Pita-Thoma, A Sene, A Santeford, M Gordon, R Lamb, Z Dong, SC Kelly, V Cavalli, J Yoshino, RS Apte
      JCI Insight, 2017;2(9):.
      Species: Mouse
      Sample Type: Aqueous humor
    3. Mitochonic Acid 5 (MA-5) Facilitates ATP Synthase Oligomerization and Cell Survival in Various Mitochondrial Diseases
      Authors: T Matsuhashi, T Sato, SI Kanno, T Suzuki, A Matsuo, Y Oba, M Kikusato, E Ogasawara, T Kudo, K Suzuki, O Ohara, H Shimbo, F Nanto, H Yamaguchi, D Saigusa, Y Mukaiyama, A Watabe, K Kikuchi, H Shima, E Mishima, Y Akiyama, Y Oikawa, HO Hsin-Jung, Y Akiyama, C Suzuki, M Uematsu, M Ogata, N Kumagai, M Toyomizu, A Hozawa, N Mano, Y Owada, S Aiba, T Yanagisawa, Y Tomioka, S Kure, S Ito, K Nakada, KI Hayashi, H Osaka, T Abe
      EBioMedicine, 2017;0(0):.
      Species: Mouse
      Sample Type: Serum
    4. Loss of Hepatic Mitochondrial Long-Chain Fatty Acid Oxidation Confers Resistance to Diet-Induced Obesity and Glucose Intolerance
      Authors: J Lee, J Choi, ES Selen Alpe, L Zhao, T Hartung, S Scafidi, RC Riddle, MJ Wolfgang
      Cell Rep, 2017;20(3):655-667.
      Species: Mouse
      Sample Type: Serum
    5. Ghrelin protects against renal damages induced by angiotensin-II via an antioxidative stress mechanism in mice.
      Authors: Fujimura K, Wakino S, Minakuchi H, Hasegawa K, Hosoya K, Komatsu M, Kaneko Y, Shinozuka K, Washida N, Kanda T, Tokuyama H, Hayashi K, Itoh H
      PLoS ONE, 2014;9(4):e94373.
      Species: Mouse
      Sample Type: Urine
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