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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    5.5 hours
  • Sensitivity
    83 pg/mL
  • Assay Range
    250.00 - 16,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Follicular Fluid,)
  • Specificity
    Natural and recombinant human Follistatin
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    Interference observed with 1 or more available related molecules.
Control Available
Product Summary
The Quantikine Human Follistatin Immunoassay is a 5.5 hour solid-phase ELISA designed to measure human Follistatin (FS288, FS300, and FS315) in follicular fluid, cell culture supernates, serum, and plasma. It contains Sf 21-expressed recombinant human Follistatin and antibodies raised against the recombinant factor. It has been shown to accurately quantitate recombinant human Follistatin. Results obtained using natural Follistatin 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 natural Follistatin.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested in separate assays to assess inter-assay precision.
Inter-Assay Precision (Precision between assays Three samples of known concentration were tested on one plate to assess intra-assay precision.
Cell Culture Supernates
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean18505363107341645485010256
Standard Deviation59136181139312584
CV%3.22.51.78.46.45.7

Serum, EDTA Plasma, Follicular Fluid
Intra-Assay Precision Inter-Assay Precision
Sample123123
n202020404040
Mean19966047119991888560411626
Standard Deviation53144239173399918
CV%2.72.429.27.17.9

Recovery

The recovery of Follistatin spiked to three different levels in samples throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 91-107
EDTA Plasma (n=5) 96 87-102
Serum (n=5) 95 87-104
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Follistatin were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Follistatin_DFN00_ELISA_97.jpg
Background: Follistatin
Follistatin (FS) was initially identified as a follicle-stimulating hormone inhibiting substance found in ovarian follicular fluid. FS is a high-affinity activin-binding protein that can act as an activin antagonist. Two alternatively spliced follistatin mRNAs exist, encoding mature FS with 288 amino acid (aa) residues (FS-288) and 315 aa residues (FS-315). Natural FS purified from porcine ovaries is primarily a carboxy-terminal truncated form of FS-315 composed of 300 aa residues.
    Related Research Areas
    Assay Procedure
    Refer to the product for complete assay procedure.

    The conjugate must remain at 2-8 °C until 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. 100 µL Assay Diluent
    4.   Add 100 µL of Assay Diluent to each well.

    5. 100 µL Standard, Control, or Sample
    6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
    7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

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

    11. 200 µL Substrate Solution
    12.   Add 200 µL Substrate Solution to each well.
    13.   For Cell Culture Supernate Samples: Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
      For Serum, Plasma, & Follicular Fluid Samples: 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.

    Showing Results 1 - 10 of 13
    Filter your results:

    Species
    1. Relationship between bone mineral density and serum osteoprotegerin in patients with chronic heart failure.
      Authors: Chen Y, Wu Y, Yang W, Wang S, Lee C, Chou N, Hsu R, Lin Y, Lin M, Ho Y, Chen M
      PLoS ONE, 2012;7(8):e44242.
      Species: Human
      Sample Type: Serum
    2. Concentration of activin A and follistatin in follicular fluid from human small antral follicles associated to gene expression of the corresponding granulosa cells.
      Authors: Jeppesen JV, Nielsen ME, Kristensen SG, Yding Andersen C
      Mol. Cell. Endocrinol., 2011;356(1):48-54.
      Species: Human
      Sample Type: Follicular Fluid
    3. BRCA1 regulates follistatin function in ovarian cancer and human ovarian surface epithelial cells.
      Authors: Karve TM, Preet A, Sneed R
      PLoS ONE, 2012;7(6):e37697.
      Species: Human
      Sample Type: Cell Culture Supernates
    4. Exercise induces a marked increase in plasma follistatin: evidence that follistatin is a contraction-induced hepatokine.
      Authors: Hansen J, Brandt C, Nielsen AR, Hojman P, Whitham M, Febbraio MA, Pedersen BK, Plomgaard P
      Endocrinology, 2011;152(1):164-71.
    5. Evaluation of systemic follistatin as an adjuvant to stimulate muscle repair and improve motor function in Pompe mice.
      Authors: Foley JW, Bercury SD, Finn P
      Mol. Ther., 2010;18(9):1584-91.
      Species: Human
      Sample Type: recombinant follistatin in mouse serum
    6. Mammalian target of rapamycin regulates miRNA-1 and follistatin in skeletal myogenesis.
      Authors: Sun Y, Ge Y, Drnevich J, Zhao Y, Band M, Chen J
      J. Cell Biol., 2010;189(7):1157-69.
      Species: Mouse
      Sample Type: Cell Culture Supernates
    7. Effect of adenovirus-mediated overexpression of follistatin and extracellular domain of activin receptor type II on gonadotropin secretion in vitro and in vivo.
      Authors: Leal AM, Takabe K, Wang L, Donaldson CJ, MacConell LA, Bilezikjian LM, Verma IM, Vale W
      Endocrinology, 2002;143(3):964-9.
      Species: Human Follistatin transgenic rat
      Sample Type: Serum
    8. Follistatin restricts bone morphogenetic protein (BMP)-2 action on the differentiation of osteoblasts in fetal rat mandibular cells.
      Authors: Abe Y, Abe T, Aida Y, Hara Y, Maeda K
      J. Bone Miner. Res., 2004;19(8):1302-7.
      Species: Rat
      Sample Type: Cell Culture Supernates
    9. The cancer gene WWOX behaves as an inhibitor of SMAD3 transcriptional activity via direct binding.
      Authors: Ferguson B, Gao X, Zelazowski M, Lee J, Jeter C, Abba M, Aldaz C
      BMC Cancer, 2013;13(0):593.
    10. The activin A-follistatin system: potent regulator of human extracellular matrix mineralization.
      Authors: Eijken M, Swagemakers S, Koedam M, Steenbergen C, Derkx P, Uitterlinden AG, van der Spek PJ, Visser JA, de Jong FH, Pols HA, van Leeuwen JP
      FASEB J., 2007;21(11):2949-60.
      Species: Human
      Sample Type: Cell Culture Supernates
    11. Gene transfer demonstrates that muscle is not a primary target for non-cell-autonomous toxicity in familial amyotrophic lateral sclerosis.
      Authors: Miller TM, Kim SH, Yamanaka K, Hester M, Umapathi P, Arnson H, Rizo L, Mendell JR, Gage FH, Cleveland DW, Kaspar BK
      Proc. Natl. Acad. Sci. U.S.A., 2006;103(51):19546-51.
      Species: Mouse
      Sample Type: Blood
    12. Adiponectin upregulates monocytic activin A but systemic levels are not altered in obesity or type 2 diabetes.
      Authors: Weigert J, Neumeier M, Wanninger J, Schober F, Sporrer D, Weber M, Schramm A, Wurm S, Stogbauer F, Filarsky M, Schaffler A, Aslanidis C, Scholmerich J, Buechler C
      Cytokine, 2009;45(2):86-91.
      Species: Human
      Sample Type: Cell Culture Supernates
    13. Differential gene expression and regulation of the bone morphogenetic protein antagonists follistatin and gremlin in normal and osteoarthritic human chondrocytes and synovial fibroblasts.
      Authors: Tardif G, Hum D, Pelletier JP, Boileau C, Ranger P, Martel-Pelletier J
      Arthritis Rheum., 2004;50(8):2521-30.
      Species: Human
      Sample Type: Cell Culture Supernates
    Expand to show all

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