Human IL-10 Antibody

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(1 Review)
    
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  • Species Reactivity
    Human
  • Specificity
    Detects human IL-10 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant Epstein-Barr virus IL-10 is observed and less than 20% cross-reactivity with recombinant mouse IL‑10, recombinant feline IL-10, recombinant porcine IL‑10, recombinant equine IL-10,  and recombinant rat IL‑10 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL-10
    Ser19-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
Product Datasheets

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Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunohistochemistry
    5-15 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 0.2-1.0 µg/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 Human IL‑10 by Western Blot. Western blot shows conditioned media of HEK293 human embryonic kidney cell line either mock transfected or transfected with human IL-10. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017).
A specific band was detected for IL‑10 at approximately 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
IL‑10 in Human Tonsil.

IL‑10 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Immunohistochemistry
IL‑10 in Human Tonsil. IL‑10 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF‑217‑NA) at 15 µg/mL overnight at
4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human IL‑10 by Simple WesternTM. Simple Western lane view shows conditioned media of HEK293 human embryonic kidney cell line either mock transfected or transfected with human IL-10, loaded at 0.2 mg/mL. A specific band was detected for IL‑10 at approximately 23 kDa (as indicated) using 10 µg/mL of Goat Anti-Human IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.
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). Proliferation elicited by Recombinant Human IL‑10 (5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-217-NA). The ND50 is typically 0.2-1.0 µg/mL.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
    Reconstitution Buffer Available
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

14 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Role of Interleukin-10 on Nasal Polypogenesis in Patients with Chronic Rhinosinusitis with Nasal Polyps
    PLoS ONE, 2016;11(9):e0161013.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  2. The BAFF receptor TACI controls IL-10 production by regulatory B cells and CLL B cells.
    Authors: Saulep-Easton D, Vincent F, Quah P, Wei A, Ting S, Croce C, Tam C, Mackay F
    Leukemia, 2016;30(1):163-72.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. Expression of Tim-1 in primary CNS lymphoma
    Cancer Med, 2016;5(11):3235-3245.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
  4. Immune suppression and resistance mediated by constitutive activation of Wnt/beta-catenin signaling in human melanoma cells.
    J. Immunol., 2012;189(5):2110-7.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  5. IL-10 expression by primary tumor cells correlates with melanoma progression from radial to vertical growth phase and development of metastatic competence.
    Authors: Itakura E, Huang RR, Wen DR, Paul E, Wunsch PH, Cochran AJ
    Mod. Pathol., 2011;24(6):801-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  6. Irritant-induced migration of Langerhans cells coincides with an IL-10-dependent switch to a macrophage-like phenotype.
    Authors: Ouwehand K, Oosterhoff D, Breetveld M
    J. Invest. Dermatol., 2011;131(2):418-25.
    Species: Human
    Sample Type: Whole Tissue
    Application: Neut
  7. Thymic stromal lymphopoietin from trophoblasts induces dendritic cell-mediated regulatory TH2 bias in the decidua during early gestation in humans.
    Authors: Guo PF, Du MR, Wu HX, Lin Y, Jin LP, Li DJ
    Blood, 2010;116(12):2061-9.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Not specified
  8. Evolving immunosuppressive microenvironment during human cervical carcinogenesis.
    Authors: Kobayashi A, Weinberg V, Darragh T, Smith-McCune K
    Mucosal Immunol, 2008;1(5):412-20.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  9. Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells.
    Authors: Liu TY, Uemura Y, Suzuki M, Narita Y, Hirata S, Ohyama H, Ishihara O, Matsushita S
    Eur. J. Immunol., 2008;38(4):1012-23.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  10. T cell-dependent survival of CD20+ and CD20- plasma cells in human secondary lymphoid tissue.
    Authors: Withers DR, Fiorini C, Fischer RT, Ettinger R, Lipsky PE, Grammer AC
    Blood, 2007;109(11):4856-64.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  11. Leukocyte-derived IL-10 reduces subepithelial fibrosis associated with chronically inhaled endotoxin.
    Authors: Garantziotis S, Brass DM, Savov J, Hollingsworth JW, McElvania-TeKippe E, Berman K, Walker JK, Schwartz DA
    Am. J. Respir. Cell Mol. Biol., 2006;35(6):662-7.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  12. Interleukin-6 and vascular endothelial growth factor release by renal cell carcinoma cells impedes lymphocyte-dendritic cell cross-talk.
    Authors: Cabillic F, Bouet-Toussaint F, Toutirais O, Rioux-Leclercq N, Fergelot P, de La Pintiere CT, Genetet N, Patard JJ, Catros-Quemener V
    Clin. Exp. Immunol., 2006;146(3):518-23.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC
  13. Enhanced apoptotic cell clearance capacity and B cell survival factor production by IL-10-activated macrophages: implications for Burkitt's lymphoma.
    Authors: Pound JD, Batth BK, Johannessen I, Wood K
    J. Immunol., 2005;174(5):3015-23.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  14. Metastable tolerance to rhesus monkey renal transplants is correlated with allograft TGF-beta 1+CD4+ T regulatory cell infiltrates.
    Authors: Torrealba JR, Katayama M, Fechner JH, Jankowska-Gan E, Kusaka S, Schultz JM, Hu H, Hamawy MM, Jonker M, Wubben J, Doxiadis G, Bontrop R, Burlingham WJ, Knechtle SJ
    J. Immunol., 2004;172(9):5753-64.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Whole Cells
    Application: Neut
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 species: Human.
We have 1 review tested in 1 application: Immunohistochemistry.

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Images Ratings Applications Species Reviewed By Date Details
Immunohistochemistry Human IL-10 Antibody AF-217-NA Array AF-217-NA
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 IHC Human Anonymous 04/13/2018
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Immunohistochemistry Human IL-10 Antibody AF-217-NA
Immunohistochemistry: Human IL-10 Antibody [AF-217-NA].

Summary

Application Immunohistochemistry
Sample Tested Adult brain
Species Human

Other Experimental Details

Other Experimental Details Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287Used at 10ug/ml.Briefly, frozen brain sections were fixed in 4% PFA (Fisher Scientific), followed by antigen retrieval using heating in acid citric buffer (Vector, Burlingame, CA, USA). Endogenous avidin-biotin was blocked for 15 min (Vector). Sections were incubated with 10% horse serum in PBS (Biosera, Boussens, France) and Fc Receptor Blocking Solution was added (Human TruStain FcX Biolegend,London, UK). Primary antibody was added overnight at 4 °C and detected with donkey anti-goat-biotin (ab6578, Abcam), followed by streptavidin-alkaline phosphatase (SA-5100, Vector) and visualised with the Vector Blue Alkaline Phosphatase Substrate Kit III (Vector).

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