Recombinant Human Interleukin-1 Receptor Antagonist

( rHuIL-1 ra )

Certificate of Analysis and Data Sheet

Description: Recombinant Human IL-1 ra produced in E.Coli is a non-glycosylated, N-terminal methionyl form of the human naturally-occurring polypeptide chain containing 153amino acids and having a molecular mass of 17000 Dalton. The rHuIL-1 ra is purified by proprietary chromatographic techniques.

Source: Escherichia Coli.

Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation & packaging: Lyophilized protein with no additives.

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

Stability: Lyophilized rHuIL-1 ra although stable at room temperature, should be stored desiccated below 0°C. Reconstituted rHuIL-1ra 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 IL-1 ra.

 Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Arg-Pro-Ser-Gly, conforming to the sequence of native human IL-1 ra.

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

Biological Activity: Our IL-1 ra is fully biologically active when compared to standard. The ED50 as determined by the dose-dependant inhibition of IL-1 stimulation of D10S cells was found to be 0.5 ng/ml.

Endotoxin: Less than 0.1 ng/µg (IEU/µg) of IL-1 ra.

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

1.     UV spectroscopy at 280 nm using the absorbency value of 0.89 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 IL-1 ra as a Reference Standard.

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

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