Recombinant Human Macrophage Inflammatory Protein-1 alpha

(rHuMIP-1a)

Certificate of Analysis and Data Sheet

       ×       Description: Recombinant Human MIP-1a produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 70 amino acids and having a molecular mass of 7820 Dalton. rHuMIP-1a 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 0.55mg/ml solution containing no additives.

        ×        Solubility: The lyophilized rHuMIP-1a is very soluble in water and most aqueous buffers below and above the           isoelectric point.

        ×        Stability: Lyophilized rHuMIP-1a, although stable at room temperature, should be stored desiccated below 0°C.   Reconstituted rHuMIP-1a 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 silver-stained SDS-PAGE .

         (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 MIP-1a.

       ×      Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be,Ala-Ser-Leu-Ala-Ala conforming to the sequence of native human MIP-1a.

       ×         Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel. 

        ×         Biological Activity: Our rHuMIP-1a is fully biologically active when compared to standard. The Activity

        is calculated by the ability of chemo-attraction of Human monocytes using 0.5-15 ng/ml.

        ×         Endotoxin: Less than 0.1 ng/µg (IEU/µg) of rHuMIP-1a.

       ×         Protein content: Protein quantitation was carried out by two independent methods:  

1.      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).

 2.  Analysis by RP-HPLC, using a calibrated solution of MIP-1a as a Reference Standard.

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