Recombinant Human Insulin Like Growth Factor-I - rHuIGF-I



Data Sheet
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Recombinant Human Insulin Like Growth Factor-I - rHuIGF-I

Description Recombinant Human IGF-I produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 70 amino acids and having a molecular mass of 7655 Dalton. rHuIGF-I is purified by proprietary chromatographic techniques.
Source Escherichia Coli.
Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation: The protein was lyophilized after extensive dialysis against 50mM acetic acid.
Solubility: It is recommended to reconstitute the lyophilized rHuIGF-I in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability: Lyophilized rHuIGF-I although stable at room temperature for 3 weeks, should be stored desiccated below -18 °C. Upon reconstitution rHuIGF-I 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 Gly-Pro-Glu-Thr-Leu. N-terminal methionine has been completely removed enzymatically.
Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity: ProSpec's rHuIGF-I is fully biologically active when compared to standards. The ED50, calculated by the dose-dependant proliferation of murine BALB\C 3T3 cells (measured by 3H-thymidine uptake) is less then 1.0 ng/ml, corresponding to a specific activity of 1x106 U/mg. For most in-vitro applications, IGF-I exerts its biological activity in the concentration range of 0.2-20 ng/ml.
Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuIGF-I .
Protein content: Protein quantitation was carried out by two independent methods: UV spectroscopy at 280 nm using the absorbency value of 0.502 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 IGF-I as a Reference Standard.