Recombinant Human Keratinocye Growth Factor - rHuKGF



Data Sheet
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Recombinant Human Keratinocye Growth Factor - rHuKGF

Description Recombinant Human KGF produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 164 amino acids and having a molecular mass of 18995 Dalton. The rHuKGF is purified by proprietary chromatographic techniques.
Source Escherichia Coli.
Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation & Packaging The protein was lyophilized from a concentrated (10mg/ml) solution with no additives.
Solubility The lyophilized rHuKGF is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=10.42).
Stability Lyophilized rHuKGF although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuKGF is best stored refrigerated at 4°C.
Purity Greater than 99.0% as determined by: Analysis by RP-HPLC (See Figure1).Anion-exchange FPLC.Analysis by reducing and non-reducing SDS-PAGE Silver Stained (See Figure 2). (Limit of acceptance:≥98.0%. 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 KGF .
Amino acid sequence The sequence of the first five N-terminal amino acids was determined and was found to be Met-Cys-Asn-Asp-Met, which agrees with the sequence of native human KGF.
Dimers and aggregates Less than 1% as determined by silver-stained SDS-PAGE gel analysis.
Biological Activity rHuKGF is fully biologically active when compared to standards. The ED50, calculated by the dose-dependant stimulation of KGF-responsive BaF3 indicator cells (measured by 3H-thymidine uptake) is 2.0 ng/ml (Limit of acceptance: 5.0 ng/ml), corresponding to a specific activity of 106 Units/mg. rHuKGF biological activity was measured by the KGF's mitogenic activity on BaF3 cells expressing the KGF receptor.
Endotoxin Less than 0.1 ng/µg (IEU/µg) of rHuKGF.
Protein content Protein quantitation was carried out by two independent methods: UV spectroscopy at 280 nm using the absorbency value of 0.9 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 KGF as a Reference Standard.
Usage This material is for research, laboratory or further manufacturing purposes.