Human FGF-basic, cct-premium, FGF-2 recombinant Protein

Synonyms

FGF2, BFGF, FGFB, HBGF-2, basic Fibroblast growth factor (bFGF), Heparin binding growth factor-2, Prostatropin

Description of Human FGF-basic, cct-premium, FGF-2

Recombinant human Fibroblast Growth Factor-basic (hFGF-b) is a 16.5 kDa protein containing 153 amino acid residues. Interestingly, hFGF-b contains no hydrophobic leader sequence previously thought to be required for cell secretion. Basic FGF bears 55% homology to acidic FGF and also seems to exist in three forms: the 154 amino-acid form and two other truncated versions of 146 and 131 amino acids lacking the N-terminal 9 and 24 residues. hFGF-b is a heparin binding growth factor which stimulates the proliferation of a wide variety of cells including mesenchymal, neuroectodermal and endothelial cells. It also exerts a potent angiogenic activity in vivo. Acidic and basic FGF compete for the binding to 125 kDa and 145 kDa receptor species. However, acidic FGF has a higher affinity for the 125 kDa species, while basic FGF has a higher affinity for the 145 kDa species. FGF receptor activation leads to the activation of MAP kinase and protein kinase C. In general FGF’s induce the proliferative response in cells derived from mesoderm and neuroectoderm. It seems that basic FGF reduces the average doubling time by shortening the G1 phase of the cell cycle. Furthermore, it has been reported to induce the release of plasminogen activator by endothelial cells. One of the most potentially significant applications of FGF-b is related to its reported ability to induce angiogenesis.

Biological Activity

The ED₅₀ of ≤ 1 ng/ml was determined for stimulation of cell proliferation with human FGF-basic (FGF-2) in HUVECs (human umbilical vein endothelial cells), corresponding to a specific activity of ≥ 1x10⁶ IU/mg. An ED₅₀ of ≤ 0.05 ng/ml was determined by stimulation of 3H-Thymidine incorporation in BALB/c 3T3 cells, corresponding to a specific activity of ≥ 2.0 x 10⁷ units/mg. Maximal effect for DNA synthesis was observed at 1.0 -2.0 ng/ml recombinant human FGF-basic.

Reconstitution

We recommend a quick spin followed by reconstitution in sterile-filtered water to a concentration of at least 0.05 mg/ml. Do not vortex. This solution can then be diluted into other aqueous buffers. For extended storage, it is recommended to further dilute in a buffer containing a carrier protein (example 0.1% BSA) and store in working aliquots at -20°C to -80°C.

Stability

The lyophilized protein is stable below -20°C until expiry date. Reconstituted human FGF-basic should be stored in working aliquots at -20°C. Please avoid repeated freeze-thaw cycles.

Amino Acid Sequence

AGSITTLPAL PEDGGSGAFP PGHFKDPKRL YCKNGGFFLR IHPDGRVDGV REKSDPHIKL QLQAEERGVV SIKGVCANRY LAMKEDGRLL ASKCVTDECF FFERLESNNY NTYRSRKYTS WYVALKRTGQ YKLGSKTGPG QKAILFLPMS AKS

SDS-PAGE and Biological Activity of FGF-basic, cct-premium

Fig. 1: SDS-PAGE analysis of recombinant human FGF-basic, cct-premium. Samples were loaded in 15% SDS-polyacrylamide gel under reducing and non-reducing condition and stained with Coomassie blue.

Fig. 2: FGF-basic, cct-premium induced proliferation of primary human umbilical vein endothelial cells (HUVEC). The cells were stimulated using recombinant human FGF-2 and the WHO standard 90/712. Values are the means (±SD) of triplicate determinations and expressed as percentage of control.

Download Datasheet