Mouse IL-27 p28/IL-30 PE-conjugated Antibody Summary
Tyr19-Pro228 (EBI3) & Phe29-Ser234 (p28)
Accession # O35228 (EBI3) & Q8K3I6 (p28)
Applications
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.
Scientific Data
Detection of IL-27 p28/IL-30 in Raw264.7 by Flow Cytometry Raw264.7 were stained with Rat Anti-Mouse IL-27 p28/IL-30 PE‑conjugated Monoclonal Antibody (Catalog # IC1834P, filled histogram) or isotype control antibody (Catalog # IC013P, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). 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 (FAB1124A). Quadrant markers were set based on control antibody staining (IC013P). View our protocol for Staining Intracellular Molecules.
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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.
Product Datasheets
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