|61977||Human Basic Fibroblast Growth Factor (hFGF basic/FGF2)||
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.
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.
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).
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.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hFGF basic. All lots are greater than 98% pure.
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.
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.
Less than 0.01 ng endotoxin/1 μg hFGF basic.
Recombinant human FGF basic (hFGF basic) Pro143-Ser288 (Accession #NP_001997) was produced in E. coli at Cell Signaling Technology.
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).
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