Intracellular Staining by Flow Cytometry
|Detection of IL‑17D in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells were stained with Human IL‑17D Monoclonal Antibody (Catalog # MAB15041, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
|IL‑17D in Human Heart. IL‑17D was detected in immersion fixed paraffin-embedded sections of human heart using 25 µg/mL Human IL‑17D Monoclonal Antibody (Catalog # MAB15041) overnight at 4 °C. Tissue was stained with the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific labeling was localized to the sarcoplasm of cardiomyocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
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). 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). Human IL-17D cDNA encodes a 202 amino acid (aa) residues protein with a putative 17 aa signal peptide (5). Human and mouse IL-17D share 78% sequence identity. Among IL-17 family members, IL-17D is most closely related to IL-17B, sharing 27% aa sequence homology (5, 6). IL-17D is expressed preferentially in skeletal muscle, heart, adipose tissue, lung, pancreas, and nervous system (1, 5). Like other IL-17 family members, IL-17D modulates immune responses indirectly by stimulating the production of myeloid growth factors and chemokines including IL-6, IL-8, and GM-CSF (5). IL-17D has also been shown to suppress the proliferation of myeloid progenitors in colony formation assays. The receptor of IL-17D has not yet been identified. However, stimulation of IL-8 production by IL-17D is mediated through the activation of nuclear factor kappa-B (5).
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