Recombinant Human Vascular Endothelial Growth Factor

 ( rHuVEGF)

Certificate of Analysis and Data Sheet

 

       ×         Description:

Recombinant Human VEGF produced in E.Coli is a double, non-glycosylated, polypeptide chain containing 165 amino acids and having a molecular mass of 38231 Dalton.

          The rHVuEGF is purified by proprietary chromatographic techniques.

 

       ×         Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

 

       ×         Formulation & Packaging:

The protein was lyophilized from a concentrated (1mg/ml) solution with no additives.

 

       ×         Solubility:

The lyophilized rHuVEGF is very soluble in water and most aqueous buffers below and above the isoelectric point.

 

       ×         Stability:

Lyophilized rHuVEGF although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuVEGF is best stored refrigerated at 4°C.

 

      ×        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.

 

(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 VEGF .

 

       ×         Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Pro-Met-Ala-Glu, which agrees with the sequence of native human VEGF.

 

       ×         Dimers and aggregates:

Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

 

       ×         Biological Activity:

Our rHuVEGF is fully biologically active when compared to standards.

  The biological activity is  calculated by the mitogenic effect of  human dermal microvascular endothelial cells using a concentrated solution of  39.0 ng/ml.

 

       ×         Endotoxin:

Less than 0.1 ng/µg (IEU/µg) of rHuVEGF.

 

       ×         Protein content:

          Protein quantitation was carried out by two independent methods:

 

1.  UV spectroscopy at 280 nm.

 

2.  Analysis by RP-HPLC, using a calibrated solution of VEGF as a Reference Standard.

 

      ×        Usage:

This material is offered for research, laboratory or further manufacturing purposes.