IL-5 Antibody [Unconjugated]

Recombinant Equine IL-5 (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 Poly-clonal Antibody (Catalog # AF2470). The ND50 is typically 0.500-6.00 µg/mL.

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

• Reactivity: Eq
• Applications: WB, B/N, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-5 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant equine IL-5
  Leu20-Gly134
  Accession # O02699
• Specificity: Detects equine IL-5 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IL5
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 10-25 ug/mL
  • Neutralization 0.500-6.00 ug/mL
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-5 Antibody [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-5 Antibody (AF2470)(1)

Publication using AF2470

• A Fettelscho, V Fettelscho, F Thoms, C Giese, M Daniel, F Olomski, J Kamarachev, K Birkmann, M Bühler, M Kummer, A Zeltins, E Marti, TM Kündig, MF Bachmann Treating insect-bite hypersensitivity in horses with active vaccination against IL-5 J. Allergy Clin. Immunol., 2018-04-04;0(0):. 2018-04-04 [PMID: 29627082] (ELISA Detection, Equine)

Bioinformatics

• Gene Symbol: IL5
• Uniprot:
  • O02699()