Detection of IL‑10 in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) either (A) stimulated to induce Th2 cells or (B) unstimulated were stained with Rabbit Anti-Human IL‑10 Monoclonal Antibody (Catalog # MAB9210) followed by Fluorescein-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0112) and Mouse Anti-Human CD4 Alexa Fluor® 700‑conjugated Monoclonal Antibody (Catalog # FAB3791N). Quadrant markers were set based on control antibody staining (Catalog # AB-105-C). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with methanol. View our protocol for Staining Intracellular Molecules.
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 Rabbit Anti-Human IL-10 Monoclonal Antibody (Catalog # MAB9210). The ND50 is typically 75-450 ng/mL.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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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