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.
Polyclonal Goat IgG
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL-10 Ser19-Asn178 Accession # P22301
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
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.
are available in the Technical Information section on our website.
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.
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.
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
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
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).
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Immunohistochemistry: Human IL-10 Antibody [AF-217-NA].
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).