Mouse IL-27 PE-conjugated Antibody
Mouse IL-27 PE-conjugated Antibody Summary
Tyr19-Pro228 (EBI3) & Phe29-Ser234 (p28)
Accession # O35228 (EBI3) & Q8K3I6 (p28)
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of IL‑27 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line was stained with Rat Anti-Mouse IL-27 PE-conjugated Monoclonal Antibody (Catalog # IC1834P, filled histogram) or isotype control antibody (Catalog # IC013P, open histogram). View our protocol for Staining Intracellular Molecules.
Detection of IL‑27 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Rat Anti-Mouse IL-27 PE-conjugated Monoclonal Antibody (Catalog # IC1834P) and Rat Anti-Mouse Integrin aM/CD11b APC-conjugated Monoclonal Antibody (Catalog # FAB1124A). Quadrant markers were set based on control antibody staining (Catalog # IC013P). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
Background: IL-27 p28/IL-30
IL-27 is a non-covalent heterodimeric molecule that belongs to the IL-6/IL-12 family of long type I cytokines. It is composed of EBI3 (EBV-induced gene 3), a 33-34 kDa glycoprotein that is related to the p40 subunit of IL-12 and IL-23, and p28, the 28-30 kDa glycoprotein that is related to the p35 chain of IL-12. The mouse EBI3 gene encodes a 228 amino acid (aa) precursor that contains an 18 aa signal peptide and 210 aa mature protein. The mature region contains two potential N-linked glycosylation sites, two fibronectin type III domains, and two pairs of conserved cysteine residues with a WSXWS-like motif that places the molecule in the hematopoietin receptor family. Although p40, the EBI3 counterpart in IL-12, is known to form homodimers, there is no evidence to date that EBI3 also homodimerizes. EBI3 is known to heterodimerize with the IL-12 p35 subunit to form IL-35. Mature mouse EBI3 shares 91% and 61% aa sequence identity to mature rat and human EBI3, respectfully. The mouse p28 gene encodes a 234 aa precursor that contains a 28 aa signal sequence and 206 aa mature region. The mature region is characterized by the presence of four alpha -helices, placing it in the IL-6 family of helical cytokines. Mature mouse p28 shares 89% and 70% aa sequence identity with mature rat and human p28, respectfully. IL-27 is expressed by monocytes, endothelial cells and dendritic cells. IL-27 binds to and signals through a heterodimeric receptor complex composed of WSX-1 (TCCR) and gp130. Evidence suggests IL-27 interacts only with WSX-1. In addition, and in mouse but not human, p28 is secreted as a monomer and appears to activate the IL-6R complex. This is consistent with the observation that p28 also heterodimerizes with CLF1, and signals through the IL-6R:gp130 complex. IL-27 has both anti- and proinflammatory properties. As an anti‑inflammatory, IL-27 seems to induce a general negative feedback program that limits T and NK-T cell activity. At the onset of infection, IL-27 induces an IL‑12 receptor on naïve CD4+ T cells, making them susceptible to subsequent IL-12 activity that generates Th1 cells at the expense of Th2 and Th17 cells. Finally, IL-27 upregulates both MHC-II and chemokine (CXCL9; CXCL10) expression on vascular endothelium, suggesting a role for IL-27 in vascular inflammation.
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