Detects canine canine IL‑5 in direct ELISAs and Western blots. In Western blots, approximately 15% cross-reactivity with recombinant bovine IL-5, recombinant equine IL-5, and recombinant feline IL-5 is observed and less than 5% cross-reactivity with recombinant rat IL-5, recombinant mouse IL-5, and recombinant human IL-5 is observed.
Measured by its ability to neutralize IL‑5-induced proliferation in the TF‑1 human erythroleukemic cell line. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The Neutralization Dose (ND50) is typically 0.75-3.0 µg/mL in the presence of 30 ng/mL Recombinant Canine IL‑5.
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.
IL‑5 in Canine PBMCs. IL‑5 was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA using Goat Anti-Canine IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1964) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by IL‑5 and Neutralization by Canine IL‑5 Antibody. Recombinant Canine IL‑5 (Catalog # 1964-CL) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Canine IL‑5 (30 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine IL‑5 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1964). The ND50 is typically 0.75-3.0 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-5 (IL-5) is a 40‑45 kDa secreted disulfide-linked homodimeric glycoprotein that plays an important role in the differentiation, growth, and function of eosinophils. It also primes basophils for histamine and leukotriene release. In mice, IL-5 also induces the proliferation, differentiation, and immunoglobulin production of B cells especially B-1 cells that constitutively express IL-5 receptor alpha. IL-5 is primarily produced by CD4+ Th2 cells. Other cell types, including mast cells, visceral smooth muscle cells, bronchial epithelium, CD16+ NK cells, eosinophils and gamma δ T cells, can also produce IL-5. Canine IL-5 is synthesized as a 134 amino acid (aa) precursor that contains a 21 aa signal sequence and a 113 aa mature segment. Mature canine IL-5 shares 62%, 66%, 85%, 84%, 58%, and 56% aa sequence identity with mature human, guinea pig, porcine, feline, mouse, and rat IL-5, respectively. The receptor for IL-5 consists of a 60 kDa ligand-binding subunit (IL-5 R alpha ) and a 120 kDa signal-transducing subunit ( beta c) (1‑7).
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