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CERTIFICATE OF ANALYSIS AND DATA SHEET

Description:
Recombinant Human MIP-1α produced in E.Coli is a single, non-glycosylated,
polypeptide chain containing 70 amino acids and having a molecular mass of 7820 Dalton. Puified by proprietary chromatographic techniques.

Source:
Escherichia Coli.

Formulation:
The protein was lyophilized from 0.55mg/ml solution containing no additives.

Solubility:
It is recommended to reconstitute the lyophilized MIP-1α in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.

Stability:
Lyophilized MIP-1 α although stable at room temperature for 3 weeks, should be stored desiccated below -18C. Upon reconstitution MIP-1α should be stored at 4C between 2-7 days and for future use below -18C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). Please prevent freeze-thaw cycles.

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 gel.

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.

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

Biological Activity:
Fully biologically active when compared to standard. The Activity is calculated by the ability of chemo-attraction of Human monocytes using 1-10 ng/ml.

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

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-1? as a Reference Standard.

 
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