Detection of IL‑5 in PMA and Ca2+ ionomycin-treated Human PBMCs by Flow Cytometry. 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.
IL‑5 in Human Peripheral Blood Lymphocytes. 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.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
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.
(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).
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Martinez-Moczygemba, M. and D. P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
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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.
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.
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