Human Follistatin Quantikine ELISA Kit

  (21 citations)
(1 Review)
    
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Assay Procedure
Citations (21)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    5.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (10 uL), Serum (100 uL), EDTA Plasma (100 uL), Follicular Fluid (10 uL)
  • Sensitivity
    83 pg/mL
  • Assay Range
    250.0 - 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
QC20 , Quantikine Immunoassay Control Group 3 - Please Inquire
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 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
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1850 5363 10734 1645 4850 10256
Standard Deviation 59 136 181 139 312 584
CV% 3.2 2.5 1.7 8.4 6.4 5.7

Serum, EDTA Plasma, Follicular Fluid
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1996 6047 11999 1888 5604 11626
Standard Deviation 53 144 239 173 399 918
CV% 2.7 2.4 2 9.2 7.1 7.9

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 [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Follistatin
Follistatin (FST) is a secreted glycoprotein that binds and neutralizes several TGF-beta superfamily proteins including Activin, BMP-6, BMP-7, and GDF-8/Myostatin. It also binds Angiogenin and regulates hematopoietic stem cell adhesion to Fibronectin. Some Follistatin binding partners will also bind Follistatin-like proteins such as FSL-3. Of three follistatin isoforms, the full-length mature follistatin (FST315) is the most abundant and the sole form in plasma, but has lower binding affinity for both activins and heparins than alternative forms. The acidic tail is missing in the splice variant FST288, which shows highest affinity, and partial in the proteolytically produced FST303, which shows intermediate affinity. Genetic deletion of follistatin in mice, or expression of only the FST288 form, is lethal perinatally due to defects of lung, skin and musculoskeletal system. Expression of only the FST315 isoform allows survival, with defects in vascularization and female fertility.
  • Entrez Gene IDs:
    10468 (Human); 14313 (Mouse)
  • Alternate Names:
    follistatin isoform FST317; Follistatin; FS; FSActivin-binding protein; FST
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.

21 Citations: Showing 1 - 10
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Species
Sample Type
  1. DNA-Mediated Gene Therapy in a Mouse Model of Limb Girdle Muscular Dystrophy 2B
    Authors: J Ma, C Pichavant, H du Bois, M Bhakta, MP Calos
    Mol Ther Methods Clin Dev, 2017;7(0):123-131.
    Species: Mouse
    Sample Type: Tissue Homogenates
  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. In vitro drug testing based on contractile activity of C2C12 cells in an epigenetic drug model
    Authors: K Ikeda, A Ito, R Imada, M Sato, Y Kawabe, M Kamihira
    Sci Rep, 2017;7(0):44570.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. Activin A and follistatin in patients with asthma. Does severity make the difference?
    Authors: Stelios Loukides
    Respirology, 2016;0(0):.
    Species: Human
    Sample Type: BAL
  5. The differential anti-tumour effects of zoledronic acid in breast cancer - evidence for a role of the activin signaling pathway.
    Authors: Wilson C, Ottewell P, Coleman R, Holen I
    BMC Cancer, 2015;15(0):55.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. Follistatin in chondrocytes: the link between TRPV4 channelopathies and skeletal malformations.
    Authors: Leddy H, McNulty A, Lee S, Rothfusz N, Gloss B, Kirby M, Hutson M, Cohn D, Guilak F, Liedtke W
    FASEB J, 2014;28(6):2525-37.
    Species: Human
    Sample Type: Cell Culture Supernates
  7. Follistatin: a novel therapeutic for the improvement of muscle regeneration.
    Authors: Yaden B, Croy J, Wang Y, Wilson J, Datta-Mannan A, Shetler P, Milner A, Bryant H, Andrews J, Dai G, Krishnan V
    J Pharmacol Exp Ther, 2014;349(2):355-71.
    Species: Mouse
    Sample Type: Tissue Homogenates
  8. Serum activin A and follistatin levels in gestational diabetes and the association of the Activin A-Follistatin system with anthropometric parameters in offspring.
    Authors: Naf S, Escote X, Ballesteros M, Yanez R, Simon-Muela I, Gil P, Albaiges G, Vendrell J, Megia A
    PLoS ONE, 2014;9(4):e92175.
    Species: Human
    Sample Type: Serum
  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.
    Species: Human
    Sample Type: Cell Culture Supernates
  10. 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
  11. 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
  12. 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.
    Species: Human
    Sample Type: Plasma
  13. 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
  14. 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
  15. 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 Protein
  16. 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
  17. 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
  18. 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: Whole Blood
  19. 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
  20. 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
  21. 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: Transgenic Rat
    Sample Type: Serum
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