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.
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.
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.
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.
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.
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).
Immune suppression and resistance mediated by constitutive activation of Wnt/beta-catenin signaling in human melanoma cells.
IL-10 expression by primary tumor cells correlates with melanoma progression from radial to vertical growth phase and development of metastatic competence.
Thymic stromal lymphopoietin from trophoblasts induces dendritic cell-mediated regulatory TH2 bias in the decidua during early gestation in humans.
Evolving immunosuppressive microenvironment during human cervical carcinogenesis.
Distinct subsets of human invariant NKT cells differentially regulate T helper responses via dendritic cells.
Am. J. Respir. Cell Mol. Biol., 2006;35(6):662-7.
Interleukin-6 and vascular endothelial growth factor release by renal cell carcinoma cells impedes lymphocyte-dendritic cell cross-talk.
Enhanced apoptotic cell clearance capacity and B cell survival factor production by IL-10-activated macrophages: implications for Burkitt's lymphoma.