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Assay Type
Solid Phase Sandwich ELISA
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Format
96-well strip plate
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Assay Length
4.5 hours
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Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (100 uL), EDTA Plasma (100 uL)
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Sensitivity
3 pg/mL
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Assay Range
10.0 - 640 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
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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.
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Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
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Interference
No significant interference observed with available related molecules.
Control Available
QC20, Quantikine Immunoassay Control Group 3 -
Product Summary
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.
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, Serum, EDTA Plasma | Intra-Assay Precision | Inter-Assay Precision |
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean | 28.8 | 104 | 234 | 32.8 | 108 | 229 |
| Standard Deviation | 2.8 | 3.2 | 7.1 | 3 | 8 | 17.3 |
| CV% | 9.7 | 3.1 | 3 | 9.1 | 7.4 | 7.6 |
Recovery
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.
Preparation and Storage
Background: FGF basic
FGF basic (FGF-2) 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 binds to FGF R1c and 2c. Its bioactivity is modulated by a number of other binding partners including heparin, Integrin alpha V beta 3, soluble FGF R1, 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 with cell surface heparan sulfate proteoglycans (HSPG) is required for the binding and activation of FGF receptors.
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