E. coli-derived human FGF basic/FGF2/bFGF protein Pro143-Ser288 Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.
>95%, by SDS-PAGE with silver staining, under reducing conditions.
<0.01 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
Measured in a cell proliferation assay using NR6R‑3T3 mouse fibroblast cells. Raines, E.W. et al. (1985) Methods Enzymol. 109:749. The ED50 for this effect is 0.1-0.6 ng/mL.
The specific activity of Recombinant Human FGF basic/FGF2 GMP is >8.0 x 105 IU/mg, which is calibrated against the human FGF basic/FGF2 WHO International Standard (NIBSC code: 90/712).
Recombinant Human FGF basic/FGF2 (145 aa) GMP Protein, CF Scientific Data Examples
GMP-grade Recombinant Human FGF basic/FGF2 (Catalog # 3718-GMP) stimulates proliferation of the NR6R‑3T3 mouse fibroblast cell line. The ED50 for this effect is 0.1-0.6 ng/mL.
1 μg/lane of Recombinant Human GMP-grade FGF basic/FGF2 was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 16 kDa.
MALDI-TOF analysis of GMP-grade Recombinant Human FGF-basic/FGF2 (Catalog # 3718-GMP). The peaks at 16312 and 16409 correspond to the measured molecular weight of the intact protein. The calculated mass is 16311 Da for Ala144-Ser288 and 16408 Da for Pro143-Ser288.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant
protein to be stored at a more dilute concentration.
The carrier free version does not contain BSA.
What formulation is right for me?
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or
as an ELISA standard.
In contrast, the carrier free protein is recommended for applications, in which the presence of BSA
Lyophilized from a 0.2 μm filtered solution in Tris-HCl and NaCl.
Reconstitute at 100 μg/mL in PBS.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, ≤ -20 °C under sterile conditions after reconstitution.
Background: FGF basic/FGF2/bFGF
FGF basic (also known as FGF2 and HBGF-2) is an 18-34 kDa, heparin-binding member of the FGF superfamily of molecules (1-3). Superfamily members are characterized by the presence of a centrally placed beta -trefoil structure. FGF acidic (FGF-1) and FGF basic (FGF2) were the first two identified FGFs, and the designations acidic and basic refer to their relative isoelectric points. Human FGF basic is 288 amino acids (aa) in length. There are multiple start sites, four of which utilize atypical CUG codons, and one that initiates at an AUG start site (4 - 6). The four CUG start sites generate high molecular weight (HMW) FGF basic. There is a 34 kDa, 288 aa form, a 24 kDa, 210 aa form, a 22.5 kDa, 201 aa form, and a 22 kDa, 196 aa form. All are retained intracellularly, undergo extensive methylation, and possess one or more nuclear localization signals (NLS) (7-9). The AUG initiating form is 18 kDa and 155 aa in length. There is no signal sequence (ss). It is, however, secreted directly through the plasma membrane via a mechanism that appears to be dependent upon tertiary structure (10). In place of a ss, there is purportedly a 9 aa N-terminal prosegment that precedes a 146 aa mature segment (11). Early isolations of 18 kDa bovine FGF basic yielded 146 aa molecules, an effect attributed to the presence of acid proteases (12). The molecule contains a heparin-binding site (aa residues 128-144), and undergoes phosphorylation at Ser117 (13). There is also an ill-defined C-terminal NLS that may be more “functional” (or 3-dimensional) than structural (7). Human 146 aa FGF basic is 97% aa identical to mouse FGF basic (14).
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SwissProt # P09038.
Klagsbrun, M. et al. (1987) Proc. Natl. Acad. Sci. USA 84:1839.