Introduction
Fibroblast growth factor-basic (FGF-basic), also known as FGF-2, is a member of the fibroblast growth factor (FGF) family. These proteins exhibit broad mitogenic and angiogenic activities, influencing various biological processes such as limb and nervous system development, wound healing, and tumor growth. FGF-basic mRNA undergoes alternative splicing and translation initiation, resulting in five isoforms with distinct properties. The isoforms initiated from a CUG codon are localized in the nucleus and contribute to intracrine effects, while the AUG-initiated isoform is primarily cytosolic, mediating paracrine and autocrine effects. FGFs act as potent mitogens for various cell types in vitro and exhibit angiogenic properties in vivo. Notably, the tissue distribution and concentration of different FGFs can vary.
Description
Recombinant Mouse Fibroblast Growth Factor-basic (FGF-2) is produced in E. coli. It is a single, non-glycosylated polypeptide chain consisting of 146 amino acids, with a molecular mass of 16.3 kDa. The protein is purified using proprietary chromatographic techniques.
Physical Appearance
Sterile white lyophilized (freeze-dried) powder.
Formulation
FGF-basic was lyophilized from a solution containing 5mM Na₂HPO₄ (pH 7.5) and 50mM NaCl.
Solubility
To reconstitute lyophilized Fibroblast Growth Factor-basic, it is recommended to dissolve the protein in sterile 18 MΩ·cm H₂O at a concentration of at least 100 µg/ml. This solution can then be further diluted into other aqueous solutions as needed.
Stability
Lyophilized Fibroblast Growth Factor-2 is stable at room temperature for up to 3 weeks; however, for long-term storage, it is recommended to store the lyophilized protein desiccated at temperatures below -18°C. After reconstitution, FGF-basic can be stored at 4°C for 2-7 days. For extended storage periods, it is advisable to store the protein at -18°C. To enhance stability during storage, consider adding a carrier protein such as 0.1% HSA or BSA. Avoid repeated freeze-thaw cycles.
Purity
The purity of the protein is greater than 95% as determined by SDS-PAGE analysis.
Biological Activity
The biological activity of FGF-basic was evaluated by measuring the dose-dependent proliferation of BALB/3T3 cells. The activity was determined to be less than 1 ng/ml, corresponding to a specific activity of 1,000,000 units/mg.
Protein Content
Protein concentration was determined using two independent methods: 1) UV spectroscopy at 280 nm, employing an absorbance value of 0.885 as the extinction coefficient for a 0.1% (1 mg/ml) solution. This value was calculated using the PC GENE computer analysis program for protein sequences (IntelliGenetics). 2) Analysis by RP-HPLC, using a calibrated solution of FGF-2 as a reference standard.
Synonyms
HBGF-2, Prostatropin, FGF-2, FGB-b.
Amino Acid Sequence
MPALPEDGGA AFPPGHFKDP KRLYCKNGGF FLRIHPDGRV DGVREKSDPH VKLQLQAEER GVVSIKGVCA NRYLAMKEDG RLLASKCVTE ECFFFERLES NNYNTYRSRK YSSWYVALKR TGQYKLGSKT GPGQKAILFL PMSAKS.
