Detection of CD25/IL-2 R alpha in Canine Blood Monocytes by Flow Cytometry.Canine peripheral blood monocytes were stained with Sheep Anti-Canine CD25/IL‑2 R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6227) followed by Phycoerythrin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0126) and Allophycocyanin-conjugated Anti-Canine CD4. Quadrant markers were set based on control antibody staining (Catalog # 5-001-A).
IL-2 receptor alpha (IL-2 R alpha ), also known as CD25, is a 55 kDa type I membrane glycoprotein that belongs to the family of cytokine receptors that utilize the common gamma chain subunit ( gamma c). IL‑2 R alpha is primarily expressed on activated T cells and on regulatory T cells (Treg) (1‑3). The canine IL-2 R alpha cDNA encodes a 268 amino acid (aa) precursor that includes a 20 aa signal peptide, a 217 aa extracellular domain (ECD) with two Sushi domains, a 19 aa transmembrane segment, and an 12 aa cytoplasmic domain (4). Within the ECD, canine IL-2 R alpha shares 49% - 60% aa sequence identity with human, mouse, and rat IL-2 R alpha. IL-2 R beta (CD122) and gamma c (IL‑2 R gamma /CD132) dimerize to form a constitutively expressed intermediate affinity IL-2 receptor (5, 6). By itself, IL-2 R alpha binds IL-2 with low affinity. It associates with IL-2 R beta and gamma c to generate a ternary high affinity IL-2 receptor complex (7). A soluble form of IL‑2 R alpha can be generated by proteolytic cleavage of the cell surface receptor, rendering the T cell unresponsive to IL-2 (8, 9). Increased serum levels of soluble IL‑2 R alpha are found in some cancers and immune disorders (10). IL-2 R alpha is required for activation induced cell death (AICD) of naive T cells, a mechanism responsible for deleting autoreactive T cell clones (11, 12). IL-2 R alpha is also required for the development of CD4+CD25+ Treg which suppress autoreactive CD4+ T cells, thereby contributing to peripheral T cell homeostasis (11‑13).
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Western Blot: Canine CD25/IL-2 R alpha Antibody [AF6227]
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