FGF Basic (FGF-2) ELISA Assay Kit


The Eagle Biosciences Human Fibroblast Growth Factor-2 (FGF Basic) ELISA Assay Kit (enzyme-linked immunoassay kit) is intended for the quantitative determination of human Fibroblast Growth Factor-2 (FGF Basic) concentrations in cell culture supernates, serum, and plasma. The Eagle Biosciences Human Fibroblast Growth Factor-2 (FGF Basic) ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: FGF31-K01 Categories: , ,

FGF Basic (FGF-2) ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 7 pg/mL
Dynamic Range: 15.625 – 500 pg/ml
Incubation Time: 3.5 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Assay Principle

The Eagle Biosciences Human Fibroblast Growth Factor-2 (FGF Basic) ELISA Assay Kit 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 colored 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.

  1. Prepare all reagents and working standards as directed in the previous sections.
  2. Add 100 µl of Standard, control, or sample, per well. Cover with the adhesive strip provided. Incubate for 1.5 hours at 37C.
  3. Aspirate each well and wash, repeating the process three times for a total of four washes. Wash by filling each well with Wash Buffer (350 µl) using a squirt bottle, manifold dispenser or auto-washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
  4. Add 100 µl of the working solution of Biotin-Conjugate to each well. Cover with a new adhesive strip and incubate 1 hour at 37C.
  5. Repeat the aspiration/wash.
  6. Add 100 µl of the working solution of Streptavidin-HRP to each well. Cover with a new adhesive strip and incubate for 30 minutes at 37C.  Avoid placing the plate in direct light.
  7. Repeat the aspiration/wash.
  8. Add 100 µl of Substrate Solution to each well. Incubate for 10-20 minutes at 37C.   Avoid placing the plate in direct light.
  9. Add 100 µl of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  10. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. (optionally 630nm as the reference wave length;610-650nm is acceptable)

Assay Background

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 (1-7). 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 (8-10). Storage pools within the cell or on cell surface heparan sulfate proteoglycans (HSPG) are likely (2). Transcription from alternate start sites produces 21-23 kDa forms found only in the nucleus (8, 9). High and low molecular weight human FGF basic isoforms target the expression of different genes (9, 10). The 18 kDa human FGF basic sequence shares 97% and 99% aa identity with mouse/rat and bovine/ovine FGF basic, respectively (6, 7). 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 (11-15). Transgenic over-expression of FGF basic mainly influences development and mineralization of bone (4, 16, 17).

Four FGF tyrosine kinase receptors (FGF R) and their splice variants show differential binding of FGFs (1). FGF basic preferentially binds FGF R1c and 2c, for which it has picomolar affinity (1, 2). 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 (2 – 4). It is upregulated in response to inflammation via mediators such as TNF-α, IL-1β, IL-2, PDGF, and nitric oxide (2). Many human tumors express FGF basic, which may correlate with tumor vascularity (2, 5).


Product Manual



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