Intracellular Staining by Flow Cytometry
|Detection of IL‑17D in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Rat Anti-Mouse IL‑17D Monoclonal Antibody (Catalog # MAB2274, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0113). 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 cysteine-knot fold near the C-terminus, but have considerable sequence divergence at the N‑terminus (1, 2, 6). 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, 6). 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 (1‑6). Mouse IL-17D is synthesized as a 205 amino acid (aa) precursor that contains a putative 24 aa signal peptide and a 181 aa mature segment. The mature region contains two potential N-linked glycosylation sites and eight cysteines, four of which are involved in the formation of a modified cysteine-knot motif (5). The molecule is reported to exist as a 53 kDa disulfide-linked homodimer (2, 5). Given that its predicted homodimeric molecular weight is 40 kDa, the molecule is presumably glycosylated. In the mature region, mouse IL-17D is 88% aa identical to human IL-17D. There is less than 30% aa identity between mouse IL-17D and other members of the mouse IL-17 family. IL-17D is expressed in skeletal muscle, adipose tissue, fetal liver, and heart, plus resting CD4+ T cells and CD19+ B cells (1). R&D Systems has shown IL-17D binding to a mouse IL-17 R/Fc construct in a functional ELISA. IL-17D is known to induce the production of IL-8, IL-6 and GM-CSF (5).
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We have 1 review tested in 1 species: Mouse.We have 1 review tested in 1 application: Western Blot.
Western Blot: Mouse IL-17D Antibody [MAB2274]