hh Human IL-6 Antibody MAB2061R-100: R&D Systems

Human IL-6 Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-6 in direct ELISAs. Does not cross-react with recombinant IL-6 from mouse, rat, or pig.
  • Source
    Recombinant Monoclonal Mouse IgG2B Clone # 1936R
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E. coli-derived recombinant human IL-6
    Val30-Met212
    Accession # P05231
  • Formulation
    Supplied as a solution in PBS. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Immunocytochemistry
    5-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑6-induced proliferation in the T1165.85.2.1 mouse plasmacytoma cell line. The Neutralization Dose (ND50) is typically 0.05-0.15 µg/mL in the presence of 2.5 ng/mL Recombinant Human IL‑6.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry
IL‑6 in Human PBMCs. IL‑6 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with LPS and monensin using Mouse Anti-Human IL‑6 Monoclonal Antibody (Catalog # MAB2061R) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Intracellular Staining by Flow Cytometry
Detection of IL‑6 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with
100 μg/mL LPS for 24 hours were stained with Mouse Anti-Human IL‑6 Monoclonal Antibody (Catalog # MAB2061R, filled histogram) or isotype control antibody (Catalog # MAB004, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Cell Proliferation Induced by IL-6 and Neutralization by Human IL-6 Antibody. Recombinant Human IL-6 (Catalog # 206-IL) stimulates proliferation in the T1165.85.2.1 mouse plasmacytoma cell line in a dose-dependent manner (orange line), as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL-6 (2.5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-6 Monoclonal Antibody (Catalog # MAB2061R). The ND50 is typically
0.05-0.15 µg/mL.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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 opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
Background: IL-6

Interleukin 6 (IL-6) is a pleiotropic alpha -helical cytokine that plays important roles in acute phase reactions, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6 activity is essential for the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. It is secreted by multiple cell types as a 22‑28 kDa phosphorylated and variably glycosylated molecule (1‑4). Mature human IL-6 is 183 amino acids (aa) in length and shares 41% aa sequence identity with mouse and rat IL-6 (5). Alternate splicing generates several isoforms with internal deletions, some of which exhibit antagonistic properties (6‑9). Human IL-6 is equally active on mouse and rat cells (10). IL-6 induces signaling through a cell surface heterodimeric receptor complex composed of a ligand binding subunit (IL-6 R) and a signal transducing subunit (gp130). IL-6 binds to IL-6 R, triggering IL-6 R association with gp130 and gp130 dimerization (11). gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM (12). Soluble forms of IL-6 R are generated by both alternate splicing and proteolytic cleavage (3). In a mechanism known as trans‑signaling, complexes of soluble IL-6 and IL-6 R elicit responses from gp130-expressing cells that lack cell surface IL-6 R (3). Trans‑signaling enables a wider range of cell types to respond to IL-6, as the expression of gp130 is ubiquitous, while that of IL-6 R is predominantly restricted to hepatocytes, leukocytes, and lymphocytes (3). Soluble splice forms of gp130 block trans‑signaling from IL-6/IL-6 R but not from other cytokines that utilize gp130 as a coreceptor (4, 13).

  • References:
    1. Van Snick, J. (1990) Annu. Rev. Immunol. 8:253.
    2. Hodge, D.R. et al. (2005) Eur. J. Cancer 41:2502.
    3. Jones, S.A. (2005) J. Immunol. 175:3468.
    4. Rose-John, S. et al. (2006) J. Leukoc. Biol. 80:227.
    5. Hirano, T. et al. (1986) Nature 324:73.
    6. Alberti, L. et al. (2005) Cancer Res. 65:2.
    7. Kestler, D.P. et al. (1995) Blood 86:4559.
    8. Kestler, D.P. et al. (1999) Am. J. Hematol. 61:169.
    9. Bihl, M.P. et al. (2002) Am. J. Respir. Cell Mol. Biol. 27:48.
    10. Chiu, C.P. et al. (1988) Proc. Natl. Acad. Sci. USA 85:7099.
    11. Murakami, M. et al. (1993) Science 260:1808.
    12. Muller-Newen, G. (2003) Sci. STKE 2003:PE40.
    13. Mitsuyama, K. et al. (2006) Clin. Exp. Immunol. 143:125. 
  • Long Name:
    Interleukin 6
  • Entrez Gene IDs:
    3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 100034196 (Equine); 493687 (Feline); 100008733 (Rabbit)
  • Alternate Names:
    B cell stimulatory factor-2; B-cell differentiation factor; BSF-2; BSF2CTL differentiation factor; CDF; HGFHSFIFNB2Hybridoma growth factor; IFN-beta-2; IL6; IL-6; IL-6B-cell stimulatory factor 2; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A
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