Intracellular Staining by Flow Cytometry
|Detection of IL‑22 in Th17 Mouse Splenocytes by Flow Cytometry. Mouse splenocytes differentiated to Th17 cells with plate-bound Rat anti-Mouse CD3 epsilon monoclonal antibody (Catalog # MAB484, 5 μg/mL) plus Goat anti-Mouse CD28 (Catalog # AF483, 5 μg/mL), Recombinant Human TGF‑ beta 1 (Catalog # 240-B, 10 ng/mL) Recombinant Mouse IL-23 (Catalog # 1887-ML, 20 ng/mL), Recombinant Mouse IL‑6 (Catalog # 406-ML, 40 ng/mL), Recombinant Mouse IL-1 beta (Catalog # 401-ML, 10 ng/mL), and Rat anti-Mouse IFN-gamma (Catalog # MAB485, 10 μg/mL) for 5 days were stained with APC-conjugated Rat anti-Mouse CD4 Monoclonal Antibody (Catalog # FAB554A) and (A) Rat Anti-Mouse IL‑22 Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC582F) or (B) isotype control antibody (Catalog # IC006F). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit. (Catalog # FC012). 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.