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 CD3 epsilon 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.
Preparation and Storage
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.
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).
Rosenberg, H. F. et al. (2007) J. Allergy Clin. Immunol. 119:1303.
Elsas, P.X. and M. I. G. Elsas (2007) Curr. Med. Chem. 14:1925.
Martinez-Moczygemba, M. and D. P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
Minamitake, Y. et al. (1990) J. Biochem. 107:292.
McKenzie, A. N. et al. (1991) Mol. Immunol. 28:155.
Shakoory, B. et al. (2004) J. Interferon Cytokine Res. 24:271.
Lalani, T. et al. (1999) Ann. Allergy Asthma Immunol. 82:317.
Sakuishi, K. et al. (2007) J. Immunol. 179:3452.
Clutterbuck, E. J. et al. (1989) Blood 73:1504.
Cameron, L. et al. (2000) J. Immunol. 164:1538.
Tavernier, J. et al. (1991) Cell 66:1175.
Zaks-Zilberman, M. et al. (2008) J. Biol. Chem. 283:13398.
Lipscombe, R. et al. (1998) J. Leukocyte Biol. 63:342.
Tavernier, J. et al. (2000) Blood 95:1600.
Kopf, M. et al. (1996) Immunity 4:15.
Horikawa, K. and K. Takatsu (2006) Immunology 118:497.
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