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.

Applications for Mouse IL-27 p28/IL-30 PE-conjugated Antibody

Application

Recommended Usage

Intracellular Staining by Flow Cytometry

10 µL/10⁶ cells
Sample: RAW 264.7 mouse monocyte/macrophage cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using IC1834P in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Formulation

Supplied in a saline solution containing BSA and Sodium Azide.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8° C. Do not use past expiration date. Protect from light.

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.

Long Name

Interleukin 27 p28

Alternate Names

IL-30, IL27 p28

Entrez Gene IDs

246778 (Human); 246779 (Mouse)

Gene Symbol

IL27

UniProt

O35228 (EBI3) & Q8K3I6 (p28)

Additional IL-27 p28/IL-30 Products

Product Documents for Mouse IL-27 p28/IL-30 PE-conjugated Antibody

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SDS

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Product Specific Notices for Mouse IL-27 p28/IL-30 PE-conjugated Antibody

For research use only

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Mouse IL-27 p28/IL-30 PE-conjugated Antibody

R&D Systems, part of Bio-Techne | Catalog # IC1834P

Key Product Details

Species Reactivity

Applications

Label

Antibody Source

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Product Specifications

Immunogen

Specificity

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Host

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Scientific Data Images for Mouse IL-27 p28/IL-30 PE-conjugated Antibody

Detection of IL-27 p28/IL-30 in Raw264.7 by Flow Cytometry