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11138600001

Roche

Interleukin-6, human (hIL-6)

recombinant (E. coli)

Quality Level

biological source

human

recombinant

expressed in E. coli

assay

>98% (SDS-PAGE)

form

solution

potency

<0.01 ng/mL EC50

mol wt

20,600 Da

packaging

pkg of 200,000 U (2 μg, 1 ml)

storage condition

avoid repeated freeze/thaw cycles

application(s)

cell culture | hybridoma: suitable

impurities

<0.1 EU/μgtested (LAL test)

UniProt accession no.

shipped in

dry ice

storage temp.

−20°C

Gene Information

human ... IL6(3569)

General description

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

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.

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.

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

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.

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).

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 hybridomas
Working 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 °C
Store the solution in aliquots at -15 to -25 °C.
Note: Avoid repeated freezing and thawing.

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.

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.

Other Notes

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

Storage Class Code

12 - Non Combustible Liquids

WGK

nwg

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

B cell growth and differentiation factors.
K Brooks et al.
Methods in enzymology, 116, 372-379 (1985-01-01)
R Bazin et al.
Journal of immunological methods, 116(2), 245-249 (1989-01-17)
The addition of macrophage feeder cells or conditioned medium has been shown to increase the yield of murine hybridomas obtained after the fusion of myeloma cells and activated B lymphocytes. It has been shown recently that the conditioned medium contains...
K Ikebuchi et al.
Proceedings of the National Academy of Sciences of the United States of America, 84(24), 9035-9039 (1987-12-01)
Interleukin-6 (IL-6, also known as B-cell stimulatory factor 2/interferon beta 2) was previously shown to support the proliferation of granulocyte/macrophage progenitors and indirectly support the formation of multilineage and blast cell colonies in cultures of spleen cells from normal mice....
Y Naitoh et al.
Biochemical and biophysical research communications, 155(3), 1459-1463 (1988-09-30)
In order to assess the effect of interleukin-6 on the hypothalamo-pituitary-adrenal axis, we administered recombinant human interleukin-6 to conscious, freely-moving rats. The intravenous injection of interleukin-6 significantly increased the plasma level of adrenocorticotropic hormone 30 min after the injection in...
R F Tiebout et al.
Journal of immunology (Baltimore, Md. : 1950), 139(10), 3402-3405 (1987-11-15)
Hybrid hybridomas are obtained by fusion of two cells, each producing its own antibody. Several authors have reported the construction of murine hybrid hybridomas with the aim to obtain bispecific monoclonal antibodies. We have investigated, in a model system, the...

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