Recombinant Ovine Prolactin

( rOPrl)

Certificate of Analysis and Data Sheet

       ×         Description:

Recombinant Ovine Prolactin produced in E.Coli is a single, non-glycosylated polypeptide chain containing 200 amino acids and having a molecular mass of 23007 Dalton.

         The rOPrl 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 (1mg/ml) solution with 0.0045mM NaHCO3.

 

       ×         Solubility:

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

 

       ×         Stability:

Lyophilized rOPrl although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rOPrl 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 Ovine Prolactin.

 

       ×         Amino acid sequence:

The sequence of the first five N-terminal amino acids was determined and was found to be Ala-Thr-Pro-Val-Cys, conforming to the sequence of native Ovine Prolactin.

 

       ×         Dimers and aggregates:

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

 

       ×         Biological Activity:

Our rOPrl is fully biologically active when compared to standard.

The ED50 as determined by the dose-dependant stimulation of the proliferation of rat lymphoma, Nb2-11 was found to be < 0.065 ng/ml.

 

       ×         Endotoxin:

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

 

       ×         Protein content:

 Protein quantitation was carried out by two independent methods:

 

         1. UV spectroscopy at 280 nm using the absorbency value of 0.85 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 rOPrl as a Reference Standard.

 

      ×         Usage:

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