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Human IL-10 Antibody

Catalog #: AF-217-NA
26 Citations 1 Review Datasheet / COA / SDS
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AF-217-NA
AF-217-SP
Detection of Human IL‑10 by Western Blot.
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Citations (26)
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Reviews (1)
Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human IL-10 Antibody Summary

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 either lyophilized or 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
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.

Scientific Data

Western Blot Detection of Human IL-10 antibody by Western Blot. View Larger

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 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

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 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

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.

Simple Western Detection of Human IL-10 antibody by Simple Western<sup>TM</sup>. View Larger

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.

Neutralization Cell Proliferation Induced by IL‑10 and Neutralization by Human IL‑10 Antibody. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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); 102133450 (Cynomolgus Monkey); 493683 (Feline); 100715618 (Guinea Pig); 2949786 (Viral)
Alternate Names
CSIF; CSIFMGC126450; Cytokine synthesis inhibitory factor; GVHDS; IL10; IL-10; IL10A; IL-10MGC126451; interleukin 10; interleukin-10; TGIF

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Citations for Human IL-10 Antibody

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.

26 Citations: Showing 1 - 10
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  1. Induction of phenotype modifying cytokines by FERMT1 mutations.
    Authors: Heinemann A, He Y, Zimina E, Boerries M, Busch H, Chmel N, Kurz T, Bruckner-Tuderman L, Has C
    Hum. Mutat., 2011-03-01;32(4):397-406.
  2. A keratinocyte-adipocyte signaling loop is reprogrammed by loss of BTG3 to augment skin carcinogenesis
    Authors: Cheng, YC;Acedera, JD;Li, YJ;Shieh, SY;
    Cell death and differentiation
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. The role of extracellular vesicle fusion with target cells in triggering systemic inflammation
    Authors: Papareddy, P;Tapken, I;Kroh, K;Varma Bhongir, RK;Rahman, M;Baumgarten, M;Cim, EI;Györffy, L;Smeds, E;Neumann, A;Veerla, S;Olinder, J;Thorlacus, H;Ryden, C;Bartakova, E;Holub, M;Herwald, H;
    Nature communications
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  4. Highly multiplexed spatial analysis identifies tissue-resident memory T cells as drivers of ulcerative and immune checkpoint inhibitor colitis
    Authors: Mick J.M. van Eijs, José J.M. Ter Linde, Matthijs J.D. Baars, Mojtaba Amini, Miangela M. Laclé, Eelco C. Brand et al.
    iScience
  5. A Novel Fusion of IL-10 Engineered to Traffic across Intestinal Epithelium to Treat Colitis
    Authors: NC Fay, BP Muthusamy, LP Nyugen, RC Desai, A Taverner, J MacKay, M Seung, T Hunter, K Liu, A Chandalia, MP Coyle, HL Kim, S Postlethwa, K Mangat, L Song, E Seto, A Alam, CV Olson, W Feng, M Saberi, TA Mahmood, RJ Mrsny
    J Immunol, 2020-11-04;0(0):.
    Species: Human
    Sample Types: Protein
    Applications: Western Blot
  6. Expression of Tim-1 in primary CNS lymphoma
    Cancer Med, 2016-10-05;5(11):3235-3245.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  7. Role of Interleukin-10 on Nasal Polypogenesis in Patients with Chronic Rhinosinusitis with Nasal Polyps
    PLoS ONE, 2016-09-01;11(9):e0161013.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  8. Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner
    Authors: Tobias Waller, Laura Kesper, Josefine Hirschfeld, Henrik Dommisch, Johanna Kölpin, Johannes Oldenburg et al.
    Infection and Immunity
  9. 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, 2015-07-03;30(1):163-72.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  10. Immune suppression and resistance mediated by constitutive activation of Wnt/beta-catenin signaling in human melanoma cells.
    J. Immunol., 2012-07-18;189(5):2110-7.
    Species: Human
    Sample Types: Whole Cells, Whole Tissue
    Applications: IHC-P, Neutralization
  11. PLTP regulates STAT3 and NF kappa B in differentiated THP1 cells and human monocyte-derived macrophages
    Authors: S. Vuletic, W. Dong, G. Wolfbauer, C. Tang, J.J. Albers
    Biochimica et Biophysica Acta (BBA) - Molecular Cell Research
  12. Effect of Fluticasone Furoate on Interleukin 6 Secretion From Adenoid Tissues in Children With Obstructive Sleep Apnea
    Authors: Rania Esteitie, Janaki Emani, Shilpy Sharma, Dana L. Suskind, Fuad M. Baroody
    Archives of Otolaryngology–Head & Neck Surgery
  13. 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., 2011-02-11;24(6):801-9.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  14. 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., 2010-11-11;131(2):418-25.
    Species: Human
    Sample Types: Whole Tissue
    Applications: Neutralization
  15. 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, 2010-06-10;116(12):2061-9.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  16. Direct and indirect impairment of human dendritic cell function by virulent Francisella tularensis Schu S4.
    Authors: Chase JC, Celli J, Bosio CM
    Infect. Immun., 2008-11-03;77(1):180-95.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  17. Evolving immunosuppressive microenvironment during human cervical carcinogenesis.
    Authors: Kobayashi A, Weinberg V, Darragh T, Smith-McCune K
    Mucosal Immunol, 2008-07-02;1(5):412-20.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  18. 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., 2008-04-01;38(4):1012-23.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  19. 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, 2007-02-13;109(11):4856-64.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  20. 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., 2006-12-01;146(3):518-23.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  21. 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., 2006-06-29;35(6):662-7.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  22. 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., 2005-03-01;174(5):3015-23.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  23. 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., 2004-05-01;172(9):5753-64.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Types: Whole Cells
    Applications: Neutralization
  24. Correction of Arginine Metabolism with Sepiapterin-the Precursor of Nitric Oxide Synthase Cofactor BH4-Induces Immunostimulatory-shift of Breast Cancer
    Authors: Zheng X, Fernando V, Sharma V et al.
    Biochem. Pharmacol.
  25. In vivo single-cell transcriptomics reveal Klebsiella pneumoniae skews lung macrophages to promote infection
    Authors: A Dumigan, O Cappa, B Morris, J Sá Pessoa, R Calderon-G, G Mills, R Lancaster, D Simpson, A Kissenpfen, JA Bengoechea
    Embo Molecular Medicine, 2022-11-07;0(0):e16888.
  26. Pomalidomide restores immune recognition of primary effusion lymphoma through upregulation of ICAM-1 and B7-2
    Authors: P Shrestha, DA Davis, HK Jaeger, A Stream, AI Aisabor, R Yarchoan
    PloS Pathogens, 2021-01-07;17(1):e1009091.

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Human IL-10 Antibody
By Stephanie Zandee on 04/13/2018
Application: IHC Sample Tested: Adult brain Species: Human

Published in https://www.ncbi.nlm.nih.gov/pubmed/28169287
Used 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).

Immunohistochemistry Human IL-10 Antibody AF-217-NA