Porcine IL-5 Antibody

Catalog #: AF3137 Datasheet

Catalog #	Availability	Size / Price	Qty
AF3137
AF3137-SP

Cell Proliferation Induced by IL‑5 and Neutralization by Porcine IL‑5 Antibody.

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Porcine IL-5 Antibody Summary

Species Reactivity

Porcine

Specificity

Detects porcine IL-5 in direct ELISAs and Western blots. In direct ELISAs, less than 30% cross‑reactivity with recombinant canine IL‑5 and less than 10% cross-reactivity with recombinant human IL-5, recombinant mouse IL-5, and recombinant rat IL-5 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant porcine IL-5
Ile20-Ser134
Accession # Q9MYM5

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 Porcine IL‑5 (Catalog # 3137-PL)

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 <5 µg/mL in the presence of 25 ng/mL Recombinant Porcine 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

Neutralization

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Cell Proliferation Induced by IL‑5 and Neutralization by Porcine IL‑5 Antibody. Recombinant Porcine IL-5 (Catalog # 3137-PL) stimulates proliferation in the TF-1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Porcine IL-5 (25 ng/mL) is neutralized (green line) by increasing concen-trations of Goat Anti-Porcine IL-5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3137). The ND₅₀ is typically <5 µg/mL.

Immunocytochemistry

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IL‑5 in Porcine PBMCs. IL-5 was detected in immersion fixed porcine peripheral blood mononuclear cells treated with calcium ionomycin and PMA using Goat Anti-Porcine IL-5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3137) 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 cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

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 secreted disulfide-linked homodimeric glycoprotein that belongs to the alpha -helical group of cytokines that includes IL-3, IL-5 and GM-CSF (1‑3). IL-5 is primarily produced by CD4⁺ Th2 cells, but eosinophils and mast cells also produce IL-5. Porcine IL-5 is synthesized as a 134 amino acid (aa) precursor that contains a 19 aa signal sequence and a 115 aa mature segment (5). Four alpha -helices and two cysteines that form interchain disulfide bonds with a second, antiparallel IL-5 molecule are conserved among species (3-5). Monomeric IL-5 is a predicted 14 kDa protein but usage of N-linked glycosylation sites may increase its molecular weight (5). Mature porcine IL-5 shares 90%, 88%, 86%, 85%, 84%, 66%, 68%, 63%, 63% and 59% aa sequence identity with mature bovine, sheep, cat, equine, canine, human, guinea pig, cotton rat, murine and rat IL-5, respectively. Recombinant porcine IL-5 induced proliferation in the human TF-1 cell line (5). The receptor for human IL-5 consists of a 60 kDa ligand-binding subunit (IL-5 R alpha ) and a 120 kDa signal-transducing subunit ( beta _c). It is suggested that dimeric IL-5 binding to IL-5 R alpha recruits beta _c, which subsequently covalently links with IL-5 R alpha. Two receptor complexes then associate to form the physiologic IL-5 receptor (6, 7). IL-5 binds proteoglycans, potentially enhancing its activity (8). Following receptor binding, IL-5 promotes the maturation, activation and migration of eosinophils, as demonstrated during asthmatic eosinophilic lung inflammation (1‑3). It also mobilizes eosinophils and CD34⁺ progenitors from marrow. It also enhances Ig release from B cells and contributes to IL-4 production. Finally, it primes basophils for histamine and leukotriene release (1, 2, 9).

References

Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
Zabeau, L. et al. (2003) Curr. Drug Targets Inflamm. Allergy 2:319.
Mertens, B. et al. (1996) Gene 176:273.
Sylvin, H. et al. (2000) Immunogenetics 51:59.
Bagley, C.J. et al. (1997) Blood 89:1471.
Ishino, T. et al. (2005) J. Biol. Chem. 280:22951.
Lipscombe, R. et al. (1998) J. Leukocyte Biol. 63:342.
Mattes, J. and P.S. Foster (2003) Curr. Drug Targets Inflamm. Allergy 2:169.

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