Intracellular Staining by Flow Cytometry
|Detection of IL‑22 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes either untreated (light orange filled histogram) or treated with PMA, Calcium Ionomycin, Recombinant Human TGF‑ beta 1 (Catalog # 240-B), and Recombinant Mouse IL‑6 (Catalog # 406-ML) (dark orange filled histogram) were stained with Rat Anti-Mouse IL‑22 APC‑conjugated Monoclonal Antibody (Catalog # IC582A) or isotype control antibody (Catalog # IC006A, 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.|
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. Mouse 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 human IL-22 and IL-10, respectively. The mouse IL-22 gene is localized to chromosome 10. Although it exists as a single copy gene in many mouse strains, the 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 STAT-1 and STAT-3 in several hepatoma cell lines and upregulate the production of acute phase proteins. IL-22 is produced by normal mouse T cells upon Con A activation. 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-22R (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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