Recombinant Human Insulin-Like Growth Factor I
Certificate of Analysis and Data Sheet
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 & Packaging: The protein was lyophilized after extensive dialysis against 50mM acetic acid.
Solubility: The lyophilized rHuIGF-I is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=7.6).
Stability: Lyophilized rHuIGF-I, although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuIGF-Iis best stored refrigerated at 4°C.
Purity: Greater than 99.0% as determined by:
(a) Analysis by RP-HPLC
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained
(Limit of acceptance:³98.0%. No more than 2% total impurities; no single impurity greater than 1%)
Amino Acid Composition:
In total agreement with the expected amino acid
composition of native
human IGF-I.
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, conforming the sequence of native human IGF-I. 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: Our 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 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:
1. 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).
2. Analysis by RP-HPLC, using a calibrated solution of IGF-I as a Reference Standard.
Usage: This material is offered for research, laboratory or further manufacturing purposes.
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