Intracellular Staining by Flow Cytometry
Detection of IL‑17B in M‑NFS‑60 Mouse Cell Line by Flow Cytometry.|
M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line was stained with Goat Anti-Mouse IL‑17B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1709, filled histogram) or isotype control antibody (Catalog # AB‑108‑C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
The Interleukin 17 (IL-17) family proteins, comprising six members (IL-17, IL-17B through IL-17F), are secreted, structurally related proteins that share a conserved cystine-knot fold near the C-terminus, but have considerable sequence divergence at the N-terminus (1, 2). With the exception of IL-17B, which exists as a non‑covalently linked dimer, all IL-17 family members are disulfide-linked dimers (3). IL-17 family proteins are pro-inflammatory cytokines that induce local cytokine production and are involved in the regulation of immune functions (1, 2). Two receptors (IL-17 R and IL-17B R), which are activated by IL-17 family members have been identified. In addition, at least three additional orphan type I transmembrane receptors with homology to IL-17 R, including IL-17 RL (IL-17 RC), IL-17 RD, and
IL‑17 RE, have also been reported (1‑4). Mouse IL-17B cDNA encodes a 180 amino acid residue (aa) protein with a putative 20 aa signal peptide (5). Mouse and human IL-17B share 88% aa sequence identity. Among IL-17 family members, mouse IL-17B is most closely related to mouse IL-17D, sharing 33% aa sequence homology. IL-17B is expressed highly in spinal cord, and at lower levels in brain, kidney, lung, small intestine, prostate, testes, pancreas, adrenal gland and trachea (5‑7). Expression of IL-17B has also been detected in chondrocytes in articular cartilage (2). IL-17B binds the IL-17B receptor but not IL-17 R and exhibits bioactivities distinct from those of IL-17 (5, 6).
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