Human IL-10 Antibody MAB9184-100: R&D Systems

Human IL-10 Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-10 in direct ELISAs and Western blots.
  • Source
    Monoclonal Mouse IgG1 Clone # 948505
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    S. frugiperda insect ovarian cell line Sf21-derived recombinant human IL-10
    Met1-Asn178
    Accession # P22301
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *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
  • Western Blot
    2 µg/mL
    See below
  • Immunocytochemistry
    5-25 µg/mL
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑10-induced proliferation in the MC/9-2 mouse mast cell line. The Neutralization Dose (ND50) is typically 15-90 ng/mL in the presence of 5 ng/mL Recombinant Human IL‑10.
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
Detection of IL‑10 by Western Blot. Western blot shows conditioned media from HEK293 human embryonic kidney cell line either mock transfected or transfected human IL-10. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human IL‑10 Monoclonal Antibody (Catalog # MAB9184) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL‑10 at approximately 17 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
IL‑10 in Human PBMCs. IL‑10 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human IL‑10 Monoclonal Antibody (Catalog # MAB9184) at 25 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.
Cell Proliferation Induced by IL‑10 and Neutralization by Human IL‑10 Antibody. Recombinant Human IL‑10 (Catalog # 217-IL) stimulates proliferation in the MC/9‑2 mouse mast cell line in a dose-dependent manner (orange line) as measured by Resazurin (Catalog # AR002). Proliferation elicited by Recombinant Human IL‑10 (5 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL‑10 Monoclonal Antibody (Catalog # MAB9184). The ND50 is typically
15‑90 ng/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. 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 reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-10

Interleukin 10, also known as cytokine synthesis inhibitory factor (CSIF), is the charter member of the IL-10 family of alpha -helical cytokines that also includes IL-19,
IL‑20, IL-22, IL-24, and IL-26/AK155 (1, 2). IL-10 is secreted by many activated hematopoietic cell types as well as hepatic stellate cells, keratinocytes, and placental cytotrophoblasts (2‑5). Mature human IL-10 shares 72%‑86% amino acid sequence identity with bovine, canine, equine, feline, mouse, ovine, porcine, and rat IL-10. Whereas human IL-10 is active on mouse cells, mouse IL-10 does not act on human cells (6, 7). IL-10 is a 178 amino acid molecule that contains two intrachain disulfide bridges and is expressed as a 36 kDa noncovalently associated homodimer (6, 8, 9). The IL-10 dimer binds to two IL-10 R alpha /IL-10 R1 chains, resulting in recruitment of two IL-10 R beta /IL-10 R2 chains and activation of a signaling cascade involving JAK1, TYK2, and STAT3 (10). IL-10 R beta does not bind IL-10 by itself but is required for signal transduction (1). IL-10 R beta also associates with IL‑20 R alpha, IL-22 R alpha, or IL-28 R alpha to form the receptor complexes for IL-22, IL-26, IL-28, and IL‑29
(11‑13). IL-10 is a critical molecule in the control of viral infections and allergic and autoimmune inflammation (14‑16). It promotes phagocytic uptake and Th2 responses but suppresses antigen presentation and Th1 proinflammatory responses (2).

  • References:
    1. Pestka, S. et al. (2004) Annu. Rev. Immunol. 22:929.
    2. O’Garra, A. and P. Vieira (2007) Nat. Rev. Immunol. 7:425.
    3. Mathurin, P. et al. (2002) Am. J. Physiol. Gastrointest. Liver Physiol. 282:G981.
    4. Grewe, M. et al. (1995) J. Invest. Dermatol. 104:3.
    5. Szony, B.J. et al. (1999) Mol. Hum. Reprod. 5:1059.
    6. Vieira, P. et al. (1991) Proc. Natl. Acad. Sci. 88:1172.
    7. Hsu, D.-H. et al. (1990) Science 250:830.
    8. Windsor, W.T. et al. (1993) Biochemistry 32:8807.
    9. Syto, R. et al. (1998) Biochemistry 37:16943.
    10. Kotenko, S.V. et al. (1997) EMBO J. 16:5894.
    11. Kotenko, S.V. et al. (2000) J. Biol. Chem. 276:2725.
    12. Hor, S. et al. (2004) J. Biol. Chem. 279:33343.
    13. Sheppard, P. et al. (2003) Nat. Immunol. 4:63.
    14. Fitzgerald, D.C. et al. (2007) Nat. Immunol. 8:1372.
    15. Wu, K. et al. (2007) Cell. Mol. Immunol. 4:269.
    16. Blackburn, S.D. and E.J. Wherry (2007) Trends Microbiol. 15:143.
  • Long Name:
    Interleukin 10
  • Entrez Gene IDs:
    3586 (Human); 16153 (Mouse); 25325 (Rat); 397106 (Porcine); 403628 (Canine); 493683 (Feline); 100135553 (Guinea Pig); 2949786 (Viral)
  • Alternate Names:
    CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF
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