Canine IL-5 Antibody

Catalog #: AF1964 Datasheet

Catalog #	Availability	Size / Price	Qty
AF1964
AF1964-SP

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Canine IL-5 Antibody Summary

Species Reactivity

Canine

Specificity

Detects canine canine IL‑5 in direct ELISAs and Western blots.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant canine IL-5
Phe20-Ser134
Accession # Q95J76

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Canine IL‑5 (Catalog # 1964-CL)

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

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.

Scientific Data

Immunocytochemistry

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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; NL001) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization

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Cell Proliferation Induced by IL‑5 and Neutralization by Canine IL‑5 Antibody. Recombinant Canine IL-5 (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 concen-trations 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

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Background: IL-5

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).

References

Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
Karlen, S. et al. (1998) Intern. Rev. Immunol. 16:227.
Takatsu, K. (1998) Cytokine Growth Factor Rev. 9:25.
Zabeau, L. et al. (2003) Curr. Drug Targets Inflamm. Allergy 2:319.
Yang, S. et al. (2001) J. Interferon Cytokine Res. 21:361.
Jans, D.A. and G. Hassan (1998) BioEssays 20:400.
Geijsen, N. et al. (2001) Cytokine Growth Factor Rev. 12:19.

Long Name

Interleukin 5

Entrez Gene IDs

3567 (Human); 16191 (Mouse); 24497 (Rat); 397409 (Porcine); 280825 (Bovine); 403790 (Canine); 493803 (Feline)

Alternate Names

BCDF mu; B-cell differentiation factor I; BCGFII; EDF; Eo-CSF; Eosinophil differentiation factor; IL5; IL-5; IL-5T-cell replacing factor; interleukin 5 (colony-stimulating factor, eosinophil); interleukin-5; TRF; TRFB cell differentiation factor I