Equine IL-5 Antibody

Catalog # Availability Size / Price Qty
AF2470
AF2470-SP
Equine IL-5 Antibody in Neutralization
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Equine IL-5 Antibody Summary

Species Reactivity
Equine
Specificity
Detects equine IL-5 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 50% cross-reactivity with recombinant bovine IL-5 and recombinant feline IL-5 is observed, 30% cross-reactivity with recombinant mouse IL-5 and recombinant canine IL-5 is observed, 20% cross-reactivity with recombinant rhesus macaque IL-5 is observed, and less than 10% cross-reactivity with recombinant rat IL-5 and recombinant porcine IL-5 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant equine IL-5
Leu20-Gly134
Accession # O02699
Formulation
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.
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 Equine IL‑5 (Catalog # 2470-EL)
Immunocytochemistry
10-25 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 0.5-2.0 µg/mL in the presence of 50 ng/mL Recombinant Equine 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.

Data Examples

Neutralization Cell Proliferation Induced by IL‑5 and Neutralization by Equine IL‑5 Antibody. View Larger

Cell Proliferation Induced by IL‑5 and Neutralization by Equine IL‑5 Antibody. Recombinant Equine IL‑5 (Catalog # 2470-EL) stimulates proliferation in the TF‑1 human erythroleukemic cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Equine IL‑5 (50 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Equine IL‑5 Antigen Affinity-purified Poly­clonal Antibody (Catalog # AF2470). The ND50 is typically 0.5-2.0 µg/mL.

Immunocytochemistry IL‑5 in Equine PBMCs. View Larger

IL‑5 in Equine PBMCs. IL‑5 was detected in immersion fixed equine peripheral blood mononuclear cells (PBMCs) treated with calcium ionomycin and PMA using Goat Anti-Equine IL‑5 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2470) 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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
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.

Background: IL-5

Interleukin-5 (IL-5) is a 40 kDa, secreted, heparin-binding, disulfide-linked homodimeric glycoprotein that belongs to the alpha -helical group of cytokines (1‑3). IL-5 is primarily produced by CD4+ Th2 cells, but other cell types such as eosinophils, endothelial cells, mast cells, visceral (airway) smooth muscle cells, bronchial epithelium, CD16+ NK cells and gamma δ T cells can also produce IL-5. Equine IL-5 is synthesized as a 134 amino acid (aa) precursor that contains a 19 aa signal sequence and a 115 aa mature segment. There are four alpha -helices, two potential N-linked glycosylation sites, and two cysteines that form interchain disulfide bonds with a second, antiparallel IL-5 molecule (3, 4). While human and mouse IL-5 have a potential NLS in their sequence, it is unclear if equine IL-5 has such a sequence. Mature horse IL-5 shares 71%, 89%, 88%, 83%, 66% and 63% aa sequence identity with mature human, bovine, feline, canine, 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). 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. This trimeric complex then associates with another trimeric complex to form the physiologic IL-5 receptor (6). Following binding, IL-5 has targeted effects. It promotes the maturation and migration of eosinophils, partially through the effects of eotaxin. It 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, 7).

References
  1. Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
  2. Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.  
  3. Zabeau, L. et al. (2003) Curr. Drug Targets Inflamm. Allergy 2:319.
  4. Vandergrifft, E.V. and D.W. Horohov (1998) GenBank Accession # O02699.
  5. Geijsen, N. et al. (2001) Cytokine Growth Factor Rev. 12:19.
  6. Bagley, C.J. et al. (1997) Blood 89:1471.
  7. 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

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