Principle of the Assay
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for FGF basic has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any FGF basic present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for FGF basic is added to the wells and binds to the combination of capture antibody- FGF basic in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A coloured product is formed in proportion to the amount of FGF basic present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven FGF basic standard dilutions and FGF basic sample concentration determined.
For Use with serum, plasma and cell culture supernatants. For Research Use Only. Not for use in diagnostic procedures.
FGF basic, also called FGF-2 (fibroblast growth factor-2) or HBGF-2 (heparin-binding growth factor-2), is the most intensively studied of the 22 mitogenic proteins of the FGF family. Family members share 35 - 60% amino acid (aa) identity, but only FGF acidic and basic lack signal peptides and are secreted by an alternate pathway.
The 18 kDa FGF basic isoform can be found in both the cytoplasm and the nucleus and is also the form that is secreted. Storage pools within the cell or on cell surface heparan sulfate proteoglycans (HSPG) are likely. Transcription from alternate start sites produces 21-23 kDa forms found only in the nucleus. High and low molecular weight human FGF basic isoforms target the expression of different genes. The 18 kDa human FGF basic sequence shares 97% and 99% aa identity with mouse/rat and bovine/ovine FGF basic, respectively. Expression of FGF basic is nearly ubiquitous. However, disruption of the mouse FGF basic gene gives relatively mild cardiovascular, skeletal, and neuronal phenotypes, suggesting compensation by other FGF family members. Transgenic over-expression of FGF basic mainly influences development and mineralization of bone.
Four FGF tyrosine kinase receptors (FGF R) and their splice variants show differential binding of FGFs. FGF basic preferentially binds FGF R1c and 2c, for which it has picomolar affinity. FGF basic also has a number of other binding partners that fine-tune FGF basic activities, according to their locations and quantities. FGF basic modulates such normal processes as angiogenesis, wound healing, tissue repair, learning and memory, and embryonic development and differentiation of heart, bone and brain. It is upregulated in response to inflammation via mediators such as TNF-α, IL-1β, IL-2, PDGF, and nitric oxide. Many human tumors express FGF basic, which may correlate with tumor vascularity.
|SYNONYMS||BFGF; FGFB; FGF-2; HBGF-2|
|Kit Components||96 Wells Quantity/Size|
|Aluminium pouches with a Microwell Plate coated with monoclonal antibody to human FGF basic (8﹡12)||1 plate|
|Human FGF basic Standard lyophilized, 1000 pg/ml upon reconstitution||2 vials|
|Concentrated Biotin-Conjugate anti-human FGF basic monoclonal antibody||2 vials|
|Streptavidin-HRP solution||2 vials|
|Standard /sample Diluent||1 bottle|
|Biotin-Conjugate antibody Diluent||1 bottle|
|Streptavidin-HRP Diluent||1 bottle|
|Wash Buffer Concentrate 20x (PBS with 1% Tween-20)||1 bottle|
|Substrate Solution||1 vial|
|Stop Solution||1 vial|
|Adhesive Films||4 pieces|
|Product data sheet||1 copy|
|Storage||Store at 2 - 8°C|
|REPEATABILITY||The coefficient of variation of both intra-assay and inter-assay were less than 10%.|
|SENSITIVITY||The minimum detectable dose was 4pg/mL.|
|ASSAY RANGE||7.8 - 500 pg/mL|
|SPECIFICITY||This assay recognizes both natural and recombinant human FGF basic. The factors listed below were prepared at 50ng/ml in Standard /sample Diluent and assayed for cross-reactivity and no significant cross-reactivity or interference was observed.|
Factors assayed for cross-reactivity
|Recombinant human||G-CSF, GM-CSF, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, LIF, EGF, TGF-β1, TNF-α, TNF-β|
|Recombinant mouse||IL-1β, IL-3, IL-4, IL-5, IL-6, IL-7, EGF, GM-CSF, TNF-α|
|Other proteins||human PDGF, porcine PDGF, human TGF-β1, porcine TGF-β1|
Data Analysis Assistance
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