Recombinant Human Neurotrophin-3

( rHuNT-3)

Certificate of Analysis and Data Sheet

 

 

       ×         Description:

Recombinant Human NT-3 produced in E.Coli is a single, non-glycosylated polypeptide chain containing 119 amino acids and having a molecular mass of 13606.29 Dalton.

         The rHuNT-3 is purified by proprietary chromatographic techniques.

 

       ×         Source:

Escherichia Coli.

 

       ×         Physical Appearance:

Sterile Filtered White lyophilized (freeze-dried) powder.

 

       ×         Formulation & packaging:

Lyophilized from a concentrated (1mg/ml) solution in water containing no additives. 

 

      Solubility:

The lyophilized rHuNT-3 is very soluble in water and most aqueous buffers below and above the isoelectric point (pI=10.69).

 

       ×         Stability:

Lyophilized rHuNT-3 although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuNT-3 is 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 NT-3.

 

       ×         Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Tyr-Ala-Glu-His-Lys, conforming to the sequence of native human NT-3.

 

       ×         Dimers and aggregates:

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

 

       ×         Biological Activity:

Our rHuNT-3 is fully biologically active when compared to standard.

The ED50 as determined by the dose-dependant induction of choline acetyl transferase in rat basal forebrain primary septal culture was found between 20-50 ng/ml.

 

       ×         Endotoxin:

Less than 0.1 ng/µg (IEU/µg) of rHuNT-3.

 

       ×         Protein content:

 Protein quantitation was carried out by two independent methods:

 

1. UV spectroscopy at 280 nm.

 

2.  Analysis by RP-HPLC, using a standard solution of NT-3 as a Reference Standard.

 

       ×         Usage:

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