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 Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC1834F, filled histogram) or isotype control antibody (Catalog # IC013F, 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.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
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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