Intracellular Staining by Flow Cytometry
|Detection of CCL11/Eotaxin in WS‑1 Human Cell Line by Flow Cytometry. WS‑1 human fetal skin fibroblast cell line treated with Recombinant Human TNF‑ alpha (Catalog # 210-TA) and Recombinant Human IL‑4 (Catalog # 204-IL)was stained with Mouse Anti-Human CCL11/Eotaxin PE‑conjugated Monoclonal Antibody (Catalog # IC3201P, filled histogram) or isotype control antibody (Catalog # IC002P, open histogram). 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.|
CCL11, also known as Eotaxin, is a secreted potent eosinophil chemoattractant that was originally purified from bronchoalveolar lavage fluid of guinea pigs sensitized by aerosol challenge with ovalbumin. Microsequencing of the purified protein revealed the guinea pig CCL11 to be a member of the beta (CC) chemokine family of inflammatory and immunoregulatory cytokines. cDNA clones for guinea pig, mouse and human CCL11 have been isolated. Human CCL11 is an O-glycosylated, 13 kDa peptide whose cDNA encodes a 97 amino acid residue precursor protein from which the amino-terminal 23 amino acid residues are cleaved to generate a 74 amino acid residue mature human CCL11. At the amino acid sequence level, mature human CCL11 is approximately 60% identical to mature mouse and guinea pig CCL11. CCL11 circulates as a monomer, homodimer and heterodimer in a complex with either CCL2 or CCL8. Human CCL11 is chemotactic for eosinophils, but not mononuclear cells or neutrophils. The CC chemokine receptors 3 and 5, plus CXCR3 have now been identified to be specific receptors for human CCL11. CCR3 has also been shown to serve as a cofactor for a restricted subset of primary HIV viruses and binding of CCL11 to CCR3 inhibits infection by the HIV isolates.
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