Human IL-5 Fluorescein-conjugated Antibody

Newer Version Available: IC605G
NEW

Discontinued Product

IC605F has been discontinued and is replaced by IC605G. IC605S. IC605T. IC605U. IC605N.

Detection of IL‑5 in Human PBMCs by Flow Cytometry.
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Human IL-5 Fluorescein-conjugated Antibody Summary

Species Reactivity
Human
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
Monoclonal Mouse IgG1 Clone # 9906
Purification
Protein A or G purified from ascites
Immunogen
S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑5
Ile20-Ser134
Accession # P05113
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Fluorescein (Excitation= 488 nm, Emission= 515-545 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
10 µL/106 cells
See below

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

Intracellular Staining by Flow Cytometry Detection of IL‑5 antibody in Human PBMCs antibody by Flow Cytometry. View Larger

Detection of IL‑5 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PMA and Calcium Ionomycin were stained with Mouse Anti-Human IL-5 Fluorescein-conjugated Monoclonal Antibody (Catalog # IC605F) and Mouse Anti-Human CD3e APC-conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # IC002F). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

Background: IL-5

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).

References
  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.
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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Citation for Human IL-5 Fluorescein-conjugated Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. IL-33 induces IL-9 production in human CD4+ T cells and basophils.
    Authors: Blom L, Poulsen BC, Jensen BM, Hansen A, Poulsen LK
    PLoS ONE, 2011-07-06;6(7):e21695.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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