Feline IL-5 Antibody Summary
Accession # O77515
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 Feline PBMCs. IL‑5 was detected in immersion fixed feline peripheral blood mononuclear cells (PBMCs) using Goat Anti-Feline IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1795) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Cell Proliferation Induced by IL‑5 and Neutralization by Feline IL‑5 Antibody. Recombinant Feline IL‑5 (Catalog # 1795-FL) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Feline IL‑5 (30 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Feline IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1795). The ND50is typically 0.5-2.0 µg/mL.
Preparation and Storage
- 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 T cell-derived factor that promotes the proliferation, differentiation and activation of eosinophils. In mice, IL-5 is also a growth and differentiation factor for B cells (1‑3). Various names previously used to describe IL-5 include: T cell replacing factor (TRF), B cell growth factor II (BCGFII), B cell differentiation factor μ (BCDF μ), eosinophil differentiation factor (EDF) and eosinophil colony-stimulating factor (Eo-CSF). Biologically active IL-5 is a disulfide-linked homodimer. As in human IL-5, the cDNA for cat IL-5 encodes a precursor protein with signal peptide that is cleaved to generate the secreted mature protein containing 115 amino acid (aa) residues. Feline IL-5 shares 70% and 59% aa sequence identity with human and mouse IL-5, respectively. IL-5 exerts its activity on target cells by binding to specific cell surface receptor complexes. The functional high-affinity receptor complex for IL-5 is composed of a ligand-binding alpha subunit that is specific for IL-5, and a non ligand-binding common beta subunit that is required for signal transduction. The common beta subunit is shared with the high-affinity receptor complexes for IL-3 and GM-CSF. In human, IL-5 R alpha subunit is primarily expressed on eosinophils and basophils. During eosinophil development, IL-5 up‑regulates the expression of IL-5 R alpha. In contrast, in mature eosinophils, the expression of IL-5 R alpha mRNA is down-regulated by IL-5, as well as by IL-3 and GM-CSF. Furthermore, IL-5 also down-modulates cell surface IL-5 R alpha via a proteinase-mediated process that releases the soluble IL-5 R alpha extracellular domain (4‑6).
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- Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
- Takatsu, K. (1998) Cytokine Growth Factor Rev. 9:25.
- Gregory, B. et al. (2003) J. Immunol. 170:5359.
- Hellman, C. et al. (2003) Clin. Exp. Immunol. 131:75.
- Liu, L.Y. et al. (2002) J. Immunol. 169:6459.
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