GDF-8/Myostatin Quantikine ELISA Kit DGDF80: R&D Systems

GDF-8/Myostatin Quantikine ELISA Kit

(9 citations)   
  • 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 (100 uL), Tissue Homogenates (20 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
  • Sensitivity
    5.32 pg/mL
  • Assay Range
    31.3 - 2,000 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma)
  • Specificity
    Natural and recombinant human, mouse, rat, canine, porcine, and equine GDF-8.
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
QC98, Quantikine Immunoassay Control Set 794 Human GDF-8/Myostatin - Please Inquire
Ancillary Reagent Kit Available
Product Summary
The Quantikine GDF-8/Myostatin Immunoassay is a 4.5 hour solid phase ELISA designed to measure GDF-8 levels in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant GDF-8 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant GDF-8 accurately. Results obtained using natural GDF-8 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural GDF-8.

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 a 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 kit components.
Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020202020
Mean17158311471464911058
Standard Deviation9.214.820.98.817.732.3
CV%5.42.51.863.63.1

Recovery

The recovery of GDF-8 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=4) 110 100-118
Serum and Plasma (n=12) 91 80-117
Tissue Homogenates (n=4) 100 90-120
Linearity
To assess the linearity of the assay, samples containing high concentrations of GDF-8 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
GDF-8/Myostatin Quantikine ELISA Kit
Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: GDF-8/Myostatin
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 8
    • Entrez Gene IDs
      2660 (Human); 17700 (Mouse);
    • Alternate Names
      GDF8; GDF-8; GDF8growth differentiation factor 8; growth/differentiation factor 8; MSTN; Myostatin;
    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. 200 µL Conjugate
    9.   Add 200 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. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    13. 50 µL Stop Solution
    14.   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.

    9 Citations: Showing 1 - 9
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    Species
    Sample Type
    1. Activin A more prominently regulates muscle mass in primates than does GDF8
      Authors: E Latres, J Mastaitis, W Fury, L Miloscio, J Trejos, J Pangilinan, H Okamoto, K Cavino, E Na, A Papatheodo, T Willer, Y Bai, J Hae Kim, A Rafique, S Jaspers, T Stitt, AJ Murphy, GD Yancopoulo, J Gromada
      Nat Commun, 2017;8(0):15153.
      Species: Human
      Sample Type: Serum
    2. Lifelong exercise, but not short-term high-intensity interval training, increases GDF11, a marker of successful aging: a�preliminary investigation
      Authors: BT Elliott, P Herbert, N Sculthorpe, FM Grace, D Stratton, LD Hayes
      Physiol Rep, 2017;5(13):.
      Species: Human
      Sample Type: Serum
    3. Serum GDF-8 levels change dynamically during controlled ovarian hyperstimulation in patients undergoing IVF/ICSI-ET
      Authors: Lanlan Fang
      Sci Rep, 2016;6(0):28036.
      Species: Human
      Sample Type: Serum
    4. Decreasing maternal myostatin programs adult offspring bone strength in a mouse model of osteogenesis imperfecta
      Proc. Natl. Acad. Sci. U.S.A., 2016;0(0):.
      Species: Mouse
      Sample Type: Serum
    5. Efficient modification of the myostatin gene in porcine somatic cells and generation of knockout piglets.
      Authors: Rao S, Fujimura T, Matsunari H, Sakuma T, Nakano K, Watanabe M, Asano Y, Kitagawa E, Yamamoto T, Nagashima H
      Mol Reprod Dev, 2016;83(1):61-70.
      Species: Porcine
      Sample Type: Tissue Homogenates
    6. Supplementation of Magnolol Attenuates Skeletal Muscle Atrophy in Bladder Cancer-Bearing Mice Undergoing Chemotherapy via Suppression of FoxO3 Activation and Induction of IGF-1.
      Authors: Chen M, Chen Y, Lee C, Hung C, Chou T
      PLoS ONE, 2015;10(11):e0143594.
      Species: Mouse
      Sample Type: Tissue Homogenates
    7. Relationship between Blood Myostatin Levels and Kidney Function:Shimane CoHRE Study.
      Authors: Yano S, Nagai A, Isomura M, Yamasaki M, Kijima T, Takeda M, Hamano T, Nabika T
      PLoS ONE, 2015;10(10):e0141035.
      Species: Human
      Sample Type: Plasma
    8. Myostatin regulates energy homeostasis in the heart and prevents heart failure.
      Authors: Biesemann N, Mendler L, Wietelmann A, Hermann S, Schafers M, Kruger M, Boettger T, Borchardt T, Braun T
      Circ Res, 2014;115(2):296-310.
      Species: Mouse
      Sample Type: Serum
    9. Preliminary investigation into a potential role for myostatin and its receptor (ActRIIB) in lean and obese horses and ponies.
      Authors: Morrison P, Bing C, Harris P, Maltin C, Grove-White D, Argo C
      PLoS ONE, 2014;9(11):e112621.
      Species: Equine
      Sample Type: Serum
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    Sample Activation Kit 1

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    Quantikine Immunoassay Control Set 794 Human GDF-8/Myostatin

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    Quantikine ELISA Wash Buffer 1

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