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Analysis Note

Specific activity/EC₅₀: >1 x 10⁸ U/mg, ﹤0.01 ng/ml (hIL-6, NIBSC, interim standard, 88/514), at least the same specific activity (EC₅₀) compared to the indicated standard is guaranteed.

Application

Recombinant IL-6, human, is a growth factor for murine and human B-cells. It can be used to replace feeder cells in the preparation of murine and human hybridomas.

Biochem/physiol Actions

Interleukin-6 participates in several pleiotropic functions in various types of cells through its specific receptor (IL-6-R). It has been suggested that interleukin-6 may participate in the interaction between neuroendocrine and immune system by activating adrenocorticotropic hormone secretion. It has also been reported that IL-6 stimulates the bonding of receptor (IL-6-R) with a non-ligand-binding membrane glycoprotein, gp130.

General description

Interleukin-6, human (hIL-6), is produced in E. coli and purified by standard chromatographic techniques.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Physical form

Solution, filtered through 0.2 µm pore size membrane.

Preparation Note

Working concentration: 10 to 100 U/ml (0.1 1 ng/ml)10 to 100 U/ml (0.1 to 1 ng/ml) A concentration of 100 U/ml (1 ng/ml) is recommended for the preparation of B-cell hybridomasWorking solution: Dilute the concentrated IL-6 solution (200,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA [or HSA (human serum albumin)] or 1 to 10% serum.Storage conditions (working solution): -15 to -25 °CStore the solution in aliquots at -15 to -25 °C.Note: Avoid repeated freezing and thawing.

Quality

Purity: 95% pure as determined by HPLC or SDS-PAGE [Endotoxin level: ﹤0.1 EU/µg (LAL-test), ﹤10 EU/ml (LAL-test).Note: 1 EU corresponds to 0.1 ng

Reconstitution

Dilute the concentrated IL-6 solution (200,000 U/ml) with PBS or culture medium containing 1 mg/ml BSA [or HSA (human serum albumin)] or 1 to 10% serum.

Sequence

Chain Length 184 AA

The primary structure of recombinant human IL-6 is identical to that of natural human IL-6 (one polypeptide chain, 184 amino acids), however, it contains an altered sequence within the first 15 amino-terminal amino acids.

Specificity

Recombinant IL-6, human is effective on mouse and human cells.IL-6, a 26 kD protein, also known as B-cell stimulatory factor 2 (BSF-2, BCDF), interferon-β₂ (IFN-β₂), hybridoma/plasmacytoma growth factor (H(P)GF), and hepatocyte-stimulating factor is produced by many cell types (e.g., T-cells, monocytes, fibroblasts, endothelial cells). IL-6 induces the terminal maturation of activated B-cells into antibody-producing cells (B-cell differentiation activity. The hepatic acute phase response is induced by IL-6 (hepatocyte-stimulating activity). It was shown that IL-6 and IL-3 act synergistically to support the proliferation of hematopoietic stem cells (hemapoietic activity), however, IL-6 does not have antiviral activity, although it has been called IFN-β₂. High affinity receptors for IL-6 are found on many cell types.

Unit Definition

EC₅₀ definition: The amount of hIL-6 that is required to support half-maximal stimulation of cell proliferation (MTT cleavage) with 7TD1 cells (1 unit equals ﹤0.01 ng).