Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) #8910

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Source

Recombinant human FGF basic (hFGF basic) Pro143-Ser288 (Accession #NP_001997) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Based on amino acid sequencing, 60-80% of recombinant hFGF basic starts at the amino-terminal Pro143 (PALPE) and has a calculated MW of 16,539. The remainder is missing Pro143 and starts at Ala144 (ALPED). DTT-reduced and non-reduced protein migrate as 17 kDa polypeptides.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hFGF basic. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant hFGF basic was determined in a NIH/3T3 cell proliferation assay. The ED50 of each lot is between 0.2 - 1.0 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hFGF basic was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hFGF basic and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of NIH/3T3 cells treated with increasing concentrations of hFGF basic was assessed. After 24 hr treatment, cells were labeled with BrdU for 4 hrs. BrdU incorporation was determined by ELISA and the OD450-OD690 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with Human Basic Fibroblast Growth Factor for 10 minutes, using Phospho-CREB (Ser133) Antibody #9191 (upper) and CREB Antibody #9192 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg hFGF basic.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 10mM DTT and 20 μg BSA per 1 μg hFGF basic. Cystines are not required for bioactivity. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 10mM DTT. Cystines are not required for bioactivity.

Background

FGF basic (FGF2) is produced by epithelial, tumor and other cell types (1). FGF basic is involved in developmental processes and regulates differentiation, proliferation, and migration (1-6). FGF basic is a critical factor for growing embryonic stem cells in culture without inducing differentiation. FGF basic has a high affinity for heparan sulfate (1,2) and binding is a step in the FGF basic activation of FGFR tyrosine kinase. There are four distinct FGF receptors and each has multiple splice variants. FGF basic binds with high affinity to many, but not all, FGFRs. Signaling cascades activated through FGF basic binding to FGFR include the ras-raf-MAPK, PLCγ/PKC, and PI3K/AKT pathways (1).

  1. Dvorak, P. and Hampl, A. (2005) Folia Histochem Cytobiol 43, 203-8.
  2. Ornitz, D.M. and Itoh, N. (2001) Genome Biol 2, REVIEWS3005.
  3. Shi, Y. et al. (2008) Crit Rev Oncol Hematol 65, 43-53.
  4. Fontijn, D. et al. (2006) Br J Cancer 94, 1627-36.
  5. Marek, L. et al. (2009) Mol Pharmacol 75, 196-207.
  6. Acevedo, V.D. et al. (2009) Cell Cycle 8, 580-8.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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