Intracellular Staining by Flow Cytometry
|Detection of IL-17 in EL-4 Mouse Cell Line by Flow Cytometry. EL-4 mouse lymphoblast cell line was treated for 16 hours with 50 ng/mL PMA then stained with Goat Anti-Mouse IL-17 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-421-NA, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.|
IL-6 Secretion Induced by IL-17 and Neutralization by Mouse IL-17 Antibody. Recombinant Mouse IL-17 (Catalog # 421-ML) stimulates IL-6 secretion in the NIH-3T3 mouse embryonic fibroblast cell line in a dose-dependent manner (orange line), as measured by the Mouse IL-6 Quantikine ELISA Kit (Catalog # M6000B). IL-6 secretion elicited by Recombinant Mouse IL-17 (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL-17 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-421-NA). The ND50 is typically|
Interleukin 17 (also known as CTLA-8) is a T cell-expressed pleiotropic cytokine that exhibits a high degree of homology to a protein encoded by the ORF13 gene of herpesvirus Saimiri. cDNA clones encoding IL-17 have been isolated from activated rat, mouse, and human T cells. Mouse IL-17 cDNA encodes a 158 amino acid (aa) residue precursor protein with a 21 amino acid residue signal peptide that is cleaved to yield the 137 aa residue mature IL-17. Both recombinant and natural IL-17 have been shown to exist as disulfide linked homodimers. At the amino acid level, mIL-17 shows 57% and 87% sequence identity with herpesvirus and rat IL-17, respectively. An IL-17 specific mouse cell surface receptor (IL-17 R) has been cloned. While the expression of IL-17 mRNA is restricted to activated alpha beta TCR+CD4-CD8-T cells, the expression of mIL-17 R mRNA has been detected in virtually all cells and tissues tested. IL-17 exhibits multiple biological activities on a variety of cells including: the induction of IL-6 and IL-8 production in fibroblasts; the enhancement of surface expression of ICAM-1 in fibroblasts; activation of NF-kappa B and costimulation of T cell proliferation.
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