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

Catalog # Availability Size / Price Qty
IC1834P
Detection of IL-27 p28/IL-30 in Raw264.7 by Flow Cytometry
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Mouse IL-27 p28/IL-30 PE-conjugated Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse IL-27 in Western blots. This antibody recognizes the p28 subunit either as part of a p28/EBI-3 heterodimer or as a free subunit.
Source
Monoclonal Rat IgG2B Clone # 234205
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Chinese hamster ovary cell line CHO-derived recombinant mouse IL‑27
Tyr19-Pro228 (EBI3) & Phe29-Ser234 (p28)
Accession # O35228 (EBI3) & Q8K3I6 (p28)
Formulation
Supplied in a saline solution containing BSA and Sodium Azide.
Label
Phycoerythrin (Excitation= 488 nm, Emission= 565-605 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
10 µL/106 cells
See below

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

Intracellular Staining by Flow Cytometry Detection of IL-27 antibody in RAW 264.7 Mouse Cell Line antibody by Flow Cytometry. View Larger

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.

Intracellular Staining by Flow Cytometry Detection of IL-27 antibody in Mouse Splenocytes antibody by Flow Cytometry. View Larger

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

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
Entrez Gene IDs
246778 (Human); 246779 (Mouse)
Alternate Names
IL27 p28; IL-27 p28; IL27;IL-27;IL27A;IL-27A;IL27p28;IL-27p28;IL30;p28; IL-30

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Mouse IL-27 PE-conjugated Antibody
By Anonymous on 09/07/2017
Application: WB Sample Tested: ATDC5 mouse chondrogenic cell line Species: Mouse