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.
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Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
R&D Systems | Catalog # DFB50
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL)
Sensitivity
3 pg/mL
Assay Range
10-640 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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Product Summary for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
The Quantikine Human FGF basic Immunoassay kit is a 4.5 hour solid phase ELISA designed to measure human FGF basic in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human FGF basic and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human FGF basic accurately. Results obtained using natural human FGF basic 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 human FGF basic.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human FGF basic. This assay cross-reacts 40% with natural Bovine FGF-basic, 0.15% with natural Bovine FGF-acidic, 17% with recombinant Bovine FGF-basic, 18% with recombinant Mouse FGF-basic and 57% with recombinant Rat FGF-basic.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence
of human FGF basic in this assay. No medical histories were available for the donors used in this
study.
ND=Non-detectable
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Serum | ND | 0 | ND |
| EDTA plasma | 13.5 | 10 | ND-14.6 |
Cell Culture Supernates:
Human peripheral blood mononuclear cells (5 x 106
cells/mL) were cultured in RPMI
supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine,
100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells were stimulated with
10 μg/mL PHA. Aliquots of the culture supernates were removed on days 1 and 5 and assayed
for levels of natural human FGF basic. All samples measured below the lowest standard,
10 pg/mL.
K562 human chronic myelogenous leukemia cells (4 x 104
cells/mL) were cultured in RPMI
supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed on
day 3, assayed for natural human FGF basic, and measured 34 pg/mL.
PC-3 human prostate cancer cells were cultured in MEM supplemented with 10% fetal bovine
serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. The cells
were stimulated with 10 μg TNF-alpha. An aliquot of the cell culture supernate was removed,
assayed for natural human FGF basic, and measured 27 pg/mL.
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, EDTA Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 28.8 | 104 | 234 | 32.8 | 108 | 229 |
| Standard Deviation | 2.8 | 3.2 | 7.1 | 3.0 | 8.0 | 17.3 |
| CV% | 9.7 | 3.1 | 3.0 | 9.1 | 7.4 | 7.6 |
Recovery for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
The recovery of FGF basic spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 108 | 94-119 |
| EDTA Plasma (n=5) | 103 | 90-124 |
| Serum (n=5) | 106 | 86-124 |
Linearity
To assess the linearity of the assay, the following samples spiked with high concentrations of FGF basic were diluted with Calibrator Diluent and then assayed.
Scientific Data Images for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
Human FGF basic ELISA Standard Curve
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
Long Name
Fibroblast Growth Factor basic
Alternate Names
bFGF, FGF-2, FGF2, HBGF-2, Prostatropin
Gene Symbol
FGF2
Additional FGF basic/FGF2/bFGF Products
Product Documents for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
For research use only
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.Related Research Areas
Citations for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit
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Sample Tested: Platelet-Rich PlasmaVerified Customer | Posted 02/24/2024
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Protocols
View specific protocols for Human FGF basic/FGF2/bFGF Quantikine ELISA Kit (DFB50):
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.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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.
100 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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