Human FGF basic/FGF2/bFGF DuoSet ELISA Summary
* 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)
Scientific Data
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- 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.
- 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.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- 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.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- 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.
29
Citations: Showing 1 - 10
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Bone Regenerative Effect of Injectable Hypoxia Preconditioned Serum-Fibrin (HPS-F) in an Ex Vivo Bone Defect Model
Authors: Jiang, J;Röper, L;Fuchs, F;Hanschen, M;Failer, S;Alageel, S;Cong, X;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Hypoxia Preconditioned Serum (HPS) Promotes Proliferation and Chondrogenic Phenotype of Chondrocytes In Vitro
Authors: Jiang, J;Altammar, J;Cong, X;Ramsauer, L;Steinbacher, V;Dornseifer, U;Schilling, AF;Machens, HG;Moog, P;
International journal of molecular sciences
Species: Human
Sample Types: Serum
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Mast Cell Tryptase Promotes Airway Remodeling by Inducing Anti-Apoptotic and Cell Growth Properties in Human Alveolar and Bronchial Epithelial Cells
Authors: Berlin, F;Mogren, S;Ly, C;Ramu, S;Hvidtfeldt, M;Uller, L;Porsbjerg, C;Andersson, CK;
Cells
Species: Human
Sample Types: Cell Culture Supernates
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In Vitro Comparison of Lymphangiogenic Potential of Hypoxia Preconditioned Serum (HPS) and Platelet-Rich Plasma (PRP)
Authors: J Jiang, X Cong, S Alageel, U Dornseifer, AF Schilling, E Hadjipanay, HG Machens, P Moog
International Journal of Molecular Sciences, 2023-01-19;24(3):.
Species: Human
Sample Types: Hypoxia Preconditioned Serum
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Fibroblast growth factor-2 bound to specific dermal fibroblast-derived extracellular vesicles is protected from degradation
Authors: I Petit, A Levy, S Estrach, CC Féral, AG Trentin, F Dingli, D Loew, J Qu, H Zhou, C Théry, C Prunier, D Aberdam, O Ferrigno
Scientific Reports, 2022-12-22;12(1):22131.
Species: Human
Sample Types: Cell Lysates
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Pro-cancerogenic effects of spontaneous and drug-induced senescence of ovarian cancer cells in vitro and in vivo: a comparative analysis
Authors: S Rutecki, P Szulc, M Paku?a, P Uruski, A Radziemski, E Naumowicz, R Moszy?ski, A Tykarski, J Miku?a-Pie, K Ksi??ek
Journal of ovarian research, 2022-07-26;15(1):87.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2022-03-09;23(6):.
Species: Human
Sample Types: Extracellular Vesicles
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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, 2021-02-17;12(1):1087.
Species: Human
Sample Types: Serum
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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, 2021-02-09;11(2):.
Species: Human
Sample Types: Cell Lysates
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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, 2020-11-01;15(0):236-242.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2020-10-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2020-03-26;66(0):105109.
Species: N/A
Sample Types: Complex Sample Type
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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, 2019-02-20;37(0):153-174.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2018-12-19;2018(0):1353085.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2018-11-20;0(0):.
Species: Human
Sample Types: Platelet Lysates
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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, 2018-06-20;13(6):e0198682.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2018-01-10;19(1):.
Species: Human
Sample Types: Plasma
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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, 2017-09-20;2017(0):2592496.
Species: Human
Sample Types: Cell Culture Supernates
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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., 2017-06-09;0(0):.
Species: Human
Sample Types: Serum
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Gelatin device for the delivery of growth factors involved in endochondral ossification
Authors: LA Ahrens, D Vonwil, J Christense, VP Shastri
PLoS ONE, 2017-04-05;12(4):e0175095.
Species: Human
Sample Types: Recombinant Protein
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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, 2013-03-19;62(1):48-51.
Species: Human
Sample Types: Cell Culture Supernates
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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, 2011-07-13;475(7355):226-30.
Species: Mouse
Sample Types: Cell Culture Supernates
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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, 2010-07-21;133(0):2232-47.
Species: Human
Sample Types: Cell Culture Supernates
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Normal ranges of angiogenesis regulatory proteins in human platelets.
Authors: Peterson JE, Zurakowski D, Italiano JE
Am. J. Hematol., 2010-07-01;85(7):487-93.
Species: Human
Sample Types: Cell Lysates
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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., 2010-05-19;24(10):3782-95.
Species: Human
Sample Types: Cell Culture Supernates
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LTBP-2 has multiple heparin/heparan sulfate binding sites.
Authors: Parsi MK, Adams JR, Whitelock J
Matrix Biol., 2010-04-09;29(5):393-401.
Species: Human
Sample Types: Recombinant Protein
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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, 2009-02-14;58(3):153-62.
Species: Human
Sample Types: Cell Culture Supernates
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Growth factor regulation of growth factors in articular chondrocytes.
Authors: Shi S, Mercer S, Eckert GJ, Trippel SB
J. Biol. Chem., 2009-01-09;284(11):6697-704.
Species: Human
Sample Types: Whole Cells
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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, 2006-02-27;94(4):524-31.
Species: Human
Sample Types: Serum
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