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Human FGF basic/FGF2/bFGF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For five or fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human FGF basic. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Data Example

Human FGF basic ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: FGF basic/FGF2/bFGF

FGF basic/FGF2/bFGF is a growth factor that functions in angiogenesis, wound healing, tissue repair, learning and memory, and the morphogenesis of heart, bone, and brain. It is upregulated in response to inflammatory stimuli and in many tumors. FGF basic/FGF2/bFGF binds to FGFR1c and 2c. Its bioactivity is modulated by a number of other binding partners including heparin, Integrin alpha V beta 3, soluble FGFR1, FGF-BP, free gangliosides, Thrombospondin, Pentraxin 3/TSG-14, Fibrinogen, alpha 2-Macroglobulin, PDGF, and CXCL4/PF4. These molecules act as cellular coreceptors or adhesion partners, extracellular matrix decoys or reservoirs, and soluble scavengers or chaperones. In particular, the interaction of FGF basic/FGF2/bFGF with cell surface heparan sulfate proteoglycans (HSPG) is required for the binding and activation of FGF receptors.

Long Name:
Fibroblast Growth Factor basic
Entrez Gene IDs:
2247 (Human); 14173 (Mouse); 281161 (Bovine); 403857 (Canine); 100033955 (Equine)
Alternate Names:
basic fibroblast growth factor bFGF; Basic fibroblast growth factor; bFGF; FGF basic; FGF2; FGF-2; FGFBprostatropin; fibroblast growth factor 2 (basic); HBGF-2; heparin-binding growth factor 2; Prostatropin

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human FGF basic/FGF2/bFGF DuoSet ELISA

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.

23 Citations: Showing 1 - 10
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  1. Therapeutic Effects of Platelet-Derived Extracellular Vesicles in a Bioengineered Tendon Disease Model
    Authors: AL Graça, RMA Domingues, I Calejo, M Gómez-Flor, ME Gomes
    International Journal of Molecular Sciences, 2022;23(6):.
    Species: Human
    Sample Types: Extracellular Vesicles
  2. A digital single-molecule nanopillar SERS platform for predicting and monitoring immune toxicities in immunotherapy
    Authors: J Li, A Wuethrich, AAI Sina, HH Cheng, Y Wang, A Behren, PN Mainwaring, M Trau
    Nature Communications, 2021;12(1):1087.
    Species: Human
    Sample Types: Serum
  3. Initial Identification of UDP-Glucose Dehydrogenase as a Prognostic Marker in Breast Cancer Patients, Which Facilitates Epirubicin Resistance and Regulates Hyaluronan Synthesis in MDA-MB-231 Cells
    Authors: DL Vitale, I Caon, A Parnigoni, I Sevic, FM Spinelli, A Icardi, A Passi, D Vigetti, L Alaniz
    Biomolecules, 2021;11(2):.
    Species: Human
    Sample Types: Cell Lysates
  4. Heparin-conjugated collagen as a potent growth factor-localizing and stabilizing scaffold for regenerative medicine
    Authors: Y Ikegami, H Mizumachi, K Yoshida, H Ijima
    Regenerative Therapy, 2020;15(0):236-242.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Regional hyperthermia enhances mesenchymal stem cell recruitment to tumor stroma: Implications for mesenchymal stem cell-based tumor therapy
    Authors: M Tutter, C Schug, KA Schmohl, S Urnauer, C Kitzberger, N Schwenk, M Petrini, C Zach, S Ziegler, P Bartenstei, WA Weber, G Multhoff, E Wagner, LH Lindner, PJ Nelson, C Spitzweg
    Mol Ther, 2020;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  6. Standing wave-assisted acoustic droplet vaporization for single and dual payload release in acoustically-responsive scaffolds
    Authors: M Aliabouzar, A Jivani, X Lu, OD Kripfgans, JB Fowlkes, ML Fabiilli
    Ultrason Sonochem, 2020;66(0):105109.
    Species: Synthetic
    Sample Types: Synthetic emulsion construct
  7. Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells
    Authors: A Islam, I Urbarova, JA Bruun, I Martinez-Z
    Eur Cell Mater, 2019;37(0):153-174.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Comparative Analysis of Different Platelet Lysates and Platelet Rich Preparations to Stimulate Tendon Cell Biology: An In Vitro Study
    Authors: F Klatte-Sch, T Schmidt, M Uckert, S Scheffler, U Kalus, M Rojewski, H Schrezenme, A Pruss, B Wildemann
    Int J Mol Sci, 2018;19(1):.
    Species: Human
    Sample Types: Plasma
  9. Crude leaf extracts of Piperaceae species downmodulate inflammatory responses by human monocytes
    Authors: AC Finato, TF Fraga-Silv, AUC Prati, AA de Souza J, BF Mazzeu, LG Felippe, RA Pinto, MA Golim, MSP Arruda, M Furlan, J Venturini
    PLoS ONE, 2018;13(6):e0198682.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Four types of human platelet lysate, including one virally inactivated by solvent-detergent, can be used to propagate Wharton Jelly mesenchymal stromal cells
    Authors: MS Chen, TJ Wang, HC Lin, B Thierry
    New biotechnology, 2018;0(0):.
    Species: Human
    Sample Types: Platelet Lysates
  11. Fibrin Glue Enhances Adipose-Derived Stromal Cell Cytokine Secretion and Survival Conferring Accelerated Diabetic Wound Healing
    Authors: U Hopfner, MM Aitzetmuel, P Ne beta bach, MS Hu, HG Machens, ZN Maan, D Duscher
    Stem Cells Int, 2018;2018(0):1353085.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. The Proangiogenic Capabilities of Malignant Ascites Generated by Aggressive Ovarian Tumors
    Authors: J Miku?a-Pie, P Uruski, S Szubert, K Maksin, R Moszy?ski, D Szpurek, A Wo?niak, S Sajdak, A Tykarski, K Ksi??ek
    Biomed Res Int, 2017;2017(0):2592496.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Gelatin device for the delivery of growth factors involved in endochondral ossification
    Authors: LA Ahrens, D Vonwil, J Christense, VP Shastri
    PLoS ONE, 2017;12(4):e0175095.
    Species: Human
    Sample Types: Recombinant Protein
  14. Altered distribution of peripheral blood dendritic cell subsets in patients with pulmonary paracoccidioidomycosis
    Authors: J Venturini, RS Cavalcante, DV Moris, M de Assis G, AD Levorato, KH Dos Reis, MSP de Arruda, RP Mendes
    Acta Trop., 2017;0(0):.
    Species: Human
    Sample Types: Serum
  15. Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures.
    Authors: David M, Kelly E, Zoellner H
    Cytokine, 2013;62(1):48-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Tumour hypoxia promotes tolerance and angiogenesis via CCL28 and T(reg) cells.
    Authors: Facciabene A, Peng X, Hagemann IS, Balint K, Barchetti A, Wang LP, Gimotty PA, Gilks CB, Lal P, Zhang L, Coukos G
    Nature, 2011;475(7355):226-30.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  17. LTBP-2 has multiple heparin/heparan sulfate binding sites.
    Authors: Parsi MK, Adams JR, Whitelock J
    Matrix Biol., 2010;29(5):393-401.
    Species: Human
    Sample Types: Recombinant Protein
  18. Soluble forms of VEGF receptor-1 and -2 promote vascular maturation via mural cell recruitment.
    Authors: Lorquet S, Berndt S, Blacher S, Gengoux E, Peulen O, Maquoi E, Noel A, Foidart JM, Munaut C, Pequeux C
    FASEB J., 2010;24(10):3782-95.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Normal ranges of angiogenesis regulatory proteins in human platelets.
    Authors: Peterson JE, Zurakowski D, Italiano JE
    Am. J. Hematol., 2010;85(7):487-93.
    Species: Human
    Sample Types: Cell Lysates
  20. Improvement in disability after alemtuzumab treatment of multiple sclerosis is associated with neuroprotective autoimmunity.
    Authors: Jones JL, Anderson JM, Phuah CL, Fox EJ, Selmaj K, Margolin D, Lake SL, Palmer J, Thompson SJ, Wilkins A, Webber DJ, Compston DA, Coles AJ
    Brain, 2010;133(0):2232-47.
    Species: Human
    Sample Types: Cell Culture Supernates
  21. Growth factor regulation of growth factors in articular chondrocytes.
    Authors: Shi S, Mercer S, Eckert GJ, Trippel SB
    J. Biol. Chem., 2009;284(11):6697-704.
    Species: Human
    Sample Types: Whole Cells
  22. Characterization of human skin-derived mesenchymal stem cell proliferation rate in different growth conditions.
    Authors: Riekstina U, Muceniece R, Cakstina I, Muiznieks I, Ancans J
    Cytotechnology, 2008;58(3):153-62.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Circulating endothelial cells and angiogenic serum factors during neoadjuvant chemotherapy of primary breast cancer.
    Authors: Furstenberger G, von Moos R, Lucas R, Thurlimann B, Senn HJ, Hamacher J, Boneberg EM
    Br. J. Cancer, 2006;94(4):524-31.
    Species: Human
    Sample Types: Serum

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Human FGF basic DuoSet ELISA
By Anonymous on 06/02/2016
Sample Tested: Polymer electrospun fibers