Recombinant Human Interleukin-6 GMP

Source Escherichia coli.

Molecular Weight Approximately 20.8 kDa, a single non-glycosylated polypeptide chain containing 183 amino acids.

AA Sequence VPPGEDSKDV AAPHRQPLTS SERIDKQIRY ILDGISALRK ETCNKSNMCE SSKEALAENN LNLPKMAEKD GCFQSGFNEE TCLVKIITGL LEFEVYLEYL QNRFESSEEQ ARAVQMSTKV LIQFLQKKAK NLDAITTPDP TTNASLLTKL QAQNQWLQDM TTHLILRSFK EFLQSSLRAL RQM

Purity > 98 % by SDS-PAGE and HPLC analyses.

Biological Activity Assay #1: Fully biologically active when compared to standard. The ED50 as determined by a cell proliferation assay using IL-6-dependent murine 7TD1 cells is less than 0.1 ng/ml, corresponding to a specific activity of > 1.0 × 107 IU/mg.

Assay #2: Fully biologically active when compared to standard. The ED50 as determined by a cell proliferation assay using IL-6-dependent murine T1165 cells is less than 0.8 ng/ml, corresponding to a specific activity of > 1.25 × 106 IU/mg.

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

Formulation Lyophilized from a 0.2 µm filtered concentrated solution in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl.

Endotoxin Less than 0.01 EU/µg of rHuIL-6 GMP as determined by LAL method.

Reconstitution We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Reconstitute in sterile distilled water or aqueous buffer containing 0.1 % BSA to a concentration of 0.1-1.0 mg/mL. Stock solutions should be apportioned into working aliquots and stored at ≤ -20 °C. Further dilutions should be made in appropriate buffered solutions.

Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.

1 month, 2 to 8 °C under sterile conditions after reconstitution.

3 months, -20 to -70 °C under sterile conditions after reconstitution.

Usage This GMP product can be for research use or further manufacturing use.

Quality statement The manufacture and testing of this product is in compliance with ICH Q7a guidelines.

Reference 1. Ferguson-Smith AC, Chen YF, Newman MS, et al. 1988. Genomics. 2:203-8.

2. van der Poll T, Keogh CV, Guirao X, et al. 1997. J Infect Dis. 176:439-44.

3. Ming JE, Cernetti C, Steinman RM, et al. 1989. J Mol Cell Immunol. 4:203-11; discussion 211-2.

4. Bastard JP, Jardel C, Delattre J, et al. 1999. Circulation. 99:2221-2.

5. Heinrich PC, Behrmann I, Muller-Newen G, et al. 1998. Biochem J. 334 ( Pt 2):297-314.

Background Interleukin-6 (IL-6) is an interleukin that in humans is encoded by the IL-6 gene and acts as both a pro-inflammatory and anti-inflammatory cytokine. It is secreted by T cells and macrophages to stimulate immune response. Furthermore, It plays an essential role in the final differentiation of B-cells into Ig-secreting cells involved in lymphocyte and monocyte differentiation. It also induces myeloma and plasmacytoma growth and induces nerve cells differentiation acts on B-cells, T-cells, hepatocytes, hematopoietic progenitor cells and cells of the CNS. The human IL-6 is a single non-glycosylated polypeptide chain containing 183 amino acids and it signals through a cell-surface type I cytokine receptor complex consisting of the ligand-binding IL-6Rα chain (CD126), and the signal- transducing component gp130 (also called CD130). The human IL-6 shares about 40% a.a. sequence identity with mouse and rat IL-6 and it is equally active on mouse and rat cells.