Intracellular Staining by Flow Cytometry
|Detection of IL‑22BP in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line was stained with Sheep Anti-Mouse IL‑22BP APC‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # IC2376A, filled histogram) or isotype control antibody (Catalog # IC016A, 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 Binding Protein (IL-22BP), also known as CRF2-10, CRF2-X, and IL-22 RA2, is a 35‑45 kDa secreted glycoprotein belonging to the Type II Cytokine Receptor Family (CRF). IL-22 signals through a receptor complex consisting of IL-22 R and IL-10 R beta. IL-10 R beta is also a component of the receptor complexes for IL-10, IL-26, IL-28, and IL-29 (1-4). IL-10 R beta is widely expressed, and other cell types express either IL-10 R alpha or IL-22 R but not both (5). The mouse IL-22BP cDNA encodes a 230 amino acid (aa) precursor that includes a 20 aa signal sequence and two fibronectin type-III domains (6). Mouse IL-22BP shares 59% and 85% aa sequence identity with human and rat IL-22BP, respectively. Among type II cytokine receptors, IL-22BP is most closely related to the extracellular domain of IL-22 R, sharing 36% aa sequence identity. Splice variants of human IL-22BP exist but have not been detected for the mouse protein (6-8). IL-22BP binds an epitope on IL-22 that is required for the interaction of IL-22 with the transmembrane IL-22 R (8-10), thereby preventing the association of IL-22 R with IL-10 R beta (10). IL-22BP blocks IL-22-induced responses including induction of MHC class I expression, Stat1 and Stat3 activation, SOCS3 gene expression,the production of reactive oxygen species (6, 9) and the modulation of the proinflammatory effects of IL-22 (11). Mouse IL-22 BP neutralizes both mouse and human IL-22 (6). IL-22BP is secreted by epithelial cells, dendritic cells, activated B cells, and activated monocytes (6, 8, 12, 13).
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