Detects mouse, rat, and cotton rat IL-10 in direct ELISAs and Western blots. In direct ELISAs, approximately 10% cross-reactivity with recombinant human IL-10, recombinant guinea pig IL-10, recombinant viral IL-10 and recombinant canine IL-10 is observed.
Polyclonal Goat IgG
E. coli-derived recombinant rat IL‑10 Ser19-Asn178 Accession # P29456
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.
Immersion fixed rat splenocytes
Measured by its ability to neutralize IL‑10-induced proliferation in the MC/9‑2 mouse mast cell line. Thompson-Snipes, L. et al. (1991) J. Exp. Med. 173:507. The Neutralization Dose (ND50) is typically 0.3-1.0 µg/mL in the presence of 40 ng/mL Recombinant Rat 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.
Detection of Recombinant Mouse and Rat IL‑10 by Western Blot. Western blot shows 25 ng of Recombinant Mouse IL‑10 (Catalog # 417-ML), Recombinant Human IL‑10 (Catalog # 217-IL), Recombinant Rat IL‑10 (Catalog # 522-RLB), and Recombinant Human IL‑26/AK155 Monomer (Catalog # 1375-IL). PVDF Membrane was probed with 0.1 µg/mL of Goat Anti-Mouse/Rat IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF519) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). 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 3.
Cell Proliferation Induced by IL‑10 and Neutralization by Rat IL‑10 Antibody.
Recombinant Rat IL‑10 (Catalog # 522-RL) stimulates proliferation in the MC/9‑2 mouse mast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑10 (40 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat IL‑10 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF519). The ND50 is typically 0.3-1.0 µg/mL.
Preparation and Storage
Reconstitute at 0.2 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, and IL-24 (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 rat IL‑10 shares 85% amino acid sequence identity with mouse and 71%‑79% amino acid sequence identity with bovine, canine, equine, feline, human, ovine, and porcine 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 (8‑10). 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-10R2 chains and activation of a signaling cascade involving JAK1, TYK2, and STAT3 (11). 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 (12‑14). IL-10 is a critical molecule in the control of viral infections and allergic and autoimmune inflammation (15‑17). It promotes phagocytic uptake and Th2 responses but suppresses antigen presentation and Th1 proinflammatory responses (2).
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