Detection of IL‑27 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cell (PBMCs) either (A) untreated (light orange filled histogram) or (B) treated with PMA and Recombinant Human IL‑2 (Catalog # 202-IL, dark orange filled histogram) were stained with Mouse Anti-Human IL‑27 Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC25261F) or isotype control antibody (Catalog # IC003F, 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 human EBI3 gene encodes a 229 amino acid (aa) precursor that
contains a 20 aa signal peptide and 209 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.However, EBI3 is known to heterodimerize
with the p35 subunit of IL-12, generating the cytokine IL-35. Human EBI3 is 61%
aa identical to mouse EBI3. The human p28 gene encodes a 243 aa precursor that
contains a 28 aa signal sequence and 215 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. Human p28 is 70% aa identical to mouse p28.
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. 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.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.