IL-5 Antibody (9906) [Unconjugated]

IL‑5 was detected in immersion fixed human peripheral blood lymphocytes using Mouse Anti-Human IL‑5 Monoclonal Antibody (Catalog # MAB605) at 5 µg/mL for 3 hours at room temperature. Cells were stained (red) and counterstained (green). Specific labeling was localized to the cytoplasm of PBMCs. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Human PBMCs, untreated (light orange filled histogram) or activated with 50 ng/mL PMA and 500 ng/mL Ca2+ ionomycin for 5 hours (dark orange filled histogram), were stained with Human IL‑5 Monoclonal Antibody (Catalog # MAB605) or isotype control antibody (Catalog # MAB002, open histogram), followed by Fluorescein-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0103B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

• Reactivity: Hu
• Applications: WB, CyTOF-ready, Flow, ICC/IF
• Clone: 9906
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

IL-5 Antibody (9906) [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑5
  Ile20-Ser134
  Accession # P05113
• Specificity: Detects human IL-5 in direct ELISAs and Western blots. In direct ELISAs and Western blots, no cross-reactivity with recombinant mouse IL‑5 is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: IL5
• Purity Statement: Protein A or G purified from ascites

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 8-25 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 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 °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.
• 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
• Reconstitution Instructions: Reconstitute at 0.5 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 (9906) [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 secreted glycoprotein that belongs to the alpha -helical group of cytokines (1 ‑ 3). Unlike other family members, it is present as a covalently linked antiparallel dimer (4, 5). The cDNA for human IL-5 encodes a signal peptide and a 115 amino acid (aa) mature protein. Mature human IL-5 shares 70%, 70%, 62%, 71%, 70% and 66%, aa sequence identity with mouse, rat, canine, equine, feline and porcine IL-5, respectively and shows cross‑reactivity with mouse IL-5. IL-5 is primarily produced by CD4+ Th2 cells, but also by activated eosinophils, mast cells, EBV-transformed B cells, Reed-Sternberg cells in Hodgkin’s disease, and IL‑2‑stimulated invariant natural killer T cells (iNKT) (1 ‑ 3, 6 ‑ 8). IL-5 increases production and mobilization of eosinophils and CD34+ progenitors from the bone marrow and causes maturation of eosinophil precursors outside the bone marrow (1, 6, 9, 10). The receptor for human IL-5, mainly expressed by eosinophils, but also found on basophils and mast cells, consists of a unique ligand-binding subunit (IL-5 R alpha ) and a shared signal‑transducing subunit, beta c (3, 6, 11). IL-5 R alpha first binds IL-5 at low affinity, then associates with preformed beta c dimers, forming a high-affinity receptor (12). IL-5 also binds proteoglycans, potentially enhancing its activity (13). Soluble forms of IL-5 R alpha   antagonize IL-5 and can be found in vivo (10, 14). In humans, IL-5 primarily affects cells of the eosinophilic lineage, and promotes their differentiation, maturation, activation, migration and survival, while in mice IL-5 also enhances Ig class switching and release from B1 cells (1 ‑ 3, 9, 10, 15, 16). IL-5 also promotes differentiation of basophils and primes them for histamine and leukotriene release (17).

1. Rosenberg, H. F. et al. (2007) J. Allergy Clin. Immunol. 119:1303.
2. Elsas, P.X. and M. I. G. Elsas (2007) Curr. Med. Chem. 14:1925.
3. Martinez-Moczygemba, M. and D. P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
4. Minamitake, Y. et al. (1990) J. Biochem. 107:292.
5. McKenzie, A. N. et al. (1991) Mol. Immunol. 28:155.
6. Shakoory, B. et al. (2004) J. Interferon Cytokine Res. 24:271.
7. Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
8. Sakuishi, K. et al. (2007) J. Immunol. 179:3452.
9. Clutterbuck, E. J. et al. (1989) Blood 73:1504.
10. Cameron, L. et al. (2000) J. Immunol. 164:1538.
11. Tavernier, J. et al. (1991) Cell 66:1175.
12. Zaks-Zilberman, M. et al. (2008) J. Biol. Chem. 283:13398.
13. Lipscombe, R. et al. (1998) J. Leukocyte Biol. 63:342.
14. Tavernier, J. et al. (2000) Blood 95:1600.
15. Kopf, M. et al. (1996) Immunity 4:15.
16. Horikawa, K. and K. Takatsu (2006) Immunology 118:497.
17. Denburg, J. A. et al. (1991) Blood 77:1462.

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 (MAB605)(5)

Publications using MAB605

• BJ Mehrara, HJ Park, RP Kataru, J Bromberg, M Coriddi, JE Baik, J Shin, C Li, MR Cavalli, EM Encarnacio, M Lee, KJ Van Zee, E Riedel, JH Dayan Pilot Study of Anti-Th2 Immunotherapy for the Treatment of Breast Cancer-Related Upper Extremity Lymphedema Biology, 2021-09-18;10(9):. 2021-09-18 [PMID: 34571811] (IHC, Human)
• S Mukherjee, R Sengupta, D Mukhopadhy, C Braun, S Mitra, S Roy, N Kanti Das, U Chatterjee, E von Stebut, M Chatterjee Impaired activation of lesional CD8+ T-cells is associated with enhanced expression of Programmed Death-1 in Indian Post Kala-azar Dermal Leishmaniasis Sci Rep, 2019-01-24;9(1):762. 2019-01-24 [PMID: 30679687] (IHC, Human)
• Daniel Sikkema Inhibition of interleukin-5 induced false positive anti-drug antibody responses against mepolizumab through the use of a competitive blocking antibody J. Immunol. Methods, 2016-11-24;0(0):. 2016-11-24 [PMID: 27889561] (Competitive Binding Assay)
• Miller M, Ramsdell J, Friedman PJ, Cho JY, Renvall M, Broide DH Computed tomographic scan-diagnosed chronic obstructive pulmonary disease-emphysema: eotaxin-1 is associated with bronchodilator response and extent of emphysema. J. Allergy Clin. Immunol., 2007-11-01;120(5):1118-25. 2007-11-01 [PMID: 17983873] (IHC-P, Human)
• Sen Y, Chunsong H, Baojun H, Linjie Z, Qun L, San J, Qiuping Z, Junyan L, Zhang X, Jinquan T Aberration of CCR7 CD8 memory T cells from patients with systemic lupus erythematosus: an inducer of T helper type 2 bias of CD4 T cells. Immunology, 2004-06-01;112(2):274-89. 2004-06-01 [PMID: 15147571] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: IL5
• Uniprot:
  • P05113()