Recombinant Human Fibroblast Growth Factor-basic - rHuFGF-b



Data Sheet
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Recombinant Human Fibroblast Growth Factor-basic - rHuFGF-b

Description Recombinant Human FGF-b (FGF-2) produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 155 amino acids and having a molecular mass of 17353 Dalton. The rHuFGF-b is purified by proprietary chromatographic techniques.
Source Escherichia Coli.
Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation: The protein was lyophilized from a concentrated (1mg/ml) sterile solution containing 5mM Tris pH=7.5 and 150mM NaCl.
Solubility: It is recommended to reconstitute the lyophilized rHuFGF-b in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability: Lyophilized rHuFGF-b although stable at room temperature for 3 weeks, should be stored desiccated below -18 °C. Upon reconstitution rHuFGF-b should be stored at 4 °C between 2-7 days and for future use below -18 °C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.
Purity: Greater than (98.0%) as determined by:(a) Analysis by RP-HPLC. (b) Anion-exchange FPLC. (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Ala-Glu-Gly-Glu.
Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity: ProSpec's rHuFGF-b is fully biologically active when compared to standards. The ED50, calculated by the dose-dependant proliferation of BAF3 cells expressing FGF receptors (measured by 3H-thymidine uptake) is less then 0.5 ng/ml, corresponding to a specific activity of (2 x106) Units/mg.
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuFGF-b.
Protein content: Protein quantitation was carried out by two independent methods: UV spectroscopy at 280 nm using the absorbency value of 0.8511 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics). Analysis by RP-HPLC, using a calibrated solution of FGF-b as a Reference Standard.
Usage This material is for research, laboratory or further manufacturing purposes.