Intracellular Staining by Flow Cytometry
Detection of IL‑22 in Human PBMCs by Flow Cytometry.Human peripheral blood mononuclear cells (PBMCs) treated with PMA, Ca++ Ionomycin, LPS, and Recombinant Human IL‑23 (Catalog # 1290-IL) were stained with Mouse Anti-Human IL‑22 PE‑conjugated Monoclonal Antibody (Catalog # IC7821P) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # IC002P). 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.
Interleukin-22 (IL-22), also known as IL-10-related T cell-derived inducible factor (IL-TIF) was initially identified as a gene induced by IL-9 in mouse T cells and mast cells. Human IL-22 cDNA encodes a 179 amino acid (aa) residue protein with a putative 33 aa signal peptide that is cleaved to generate a 147 aa mature protein that shares approximately 79% and 22% aa sequence identity with mouse IL-22 and human IL-10, respectively. The human IL-22 gene is localized to chromosome 12q15. Although it exists as a single copy gene in human and in many mouse strains, the mouse IL-22 gene is duplicated in some mouse strains including C57B1/6, FVB and 129. The two mouse genes designated IL-TIF alpha and IL-TIF beta, share greater than 98% sequence homology in their coding region. IL-22 has been shown to activate STAT1 and STAT3 in several hepatoma cell lines and upregulate the production of acute phase proteins. IL-22 is produced by normal T cells upon anti-CD3 stimulation in humans. Mouse IL-22 expression is also induced in various organs upon lipopolysaccharide injection, suggesting that IL-22 may be involved in inflammatory responses. The functional IL-22 receptor complex consists of two receptor subunits, IL-22 R (previously an orphan receptor named CRF2-9) and IL-10R beta (previously known as CRF2-4), belonging to the class II cytokine receptor family.
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