Human IL-27 Antibody

Catalog #: MAB25261
1 Citation Datasheet / COA / SDS

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MAB25261
MAB25261-SP

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Detection of IL‑27 in PHA + IL‑2-treated Human PBMC by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human IL-27 Antibody Summary

Species Reactivity

Human

Specificity

Detects human IL-27 in direct ELISAs.

Source

Monoclonal Mouse IgG_2A Clone # 307426

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human IL‑27
Arg21-Lys229 of EBI-3 (Accession #Q14213.2) and Phe29-Pro243 of p28 (Accession #AAM34498)

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

2.5 µg/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 PHA + IL-2-treated Human PBMC antibody by Flow Cytometry.

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Detection of IL‑27 in PHA + IL‑2-treated Human PBMC by Flow Cytometry. Human PBMC, treated with 5 µg/mL PHA and 5 ng/mL rhIL-2 (dark orange filled histogram), or untreated (open histogram, yellow line) were stained with Mouse Anti-Human IL-27 Monoclonal Antibody (Catalog # MAB25261) or isotype control antibody (Catalog # MAB003, blue open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-27

IL-27 is a heterodimeric group 2 receptor ligand molecule that belongs to the IL-6/IL-12 family of long type I cytokines (1). It is composed of EBI3 (EBV-induced gene 3), a 34 kDa glycoprotein that is related to the p40 subunit of IL-12 and IL-23, and p28, the 28 kDa glycoprotein that is related to the p35 chain of IL-12 (2-4). The human EBI3 gene encodes a 229 amino acid (aa) precursor that contains a 20 aa signal peptide and 209 aa mature protein (5). 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 (5). Although p40, the EBI3 counterpart in IL-12, is known to form homodimers, there is no evidence to date that EBI3 also homodimerizes. 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 (6). 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 74% aa identical to mouse p28. IL-27 is expressed by monocytes, endothelial cells and dendritic cells (7). 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 (6, 8, 9). 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 (3, 7). 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 (and possible Th1 development) (10).

References

Boulay, J-L. et al. (2003) Immunity 19:159.
Trinchieri, G. et al. (2003) Immunity 19:641.
Murakami, M. et al. (2004) Growth Factors 22:75.
Cordoba-Rodriguez, R. and D.M. Frucht (2003) Exp. Opin. Biol. Ther. 3:715.
Devergne, O. et al. (1996) J. Virology 70:1143.
Pflanz, S. et al. (2002) Immunity 16:779.
Villarino, A.V. et al. (2004) J. Immunol. 173:715.
Pflanz, S. et al. (2004) J Immunol 172:2225.
Scheller, J. et al. (2005) Biochem. Biophys. Res. Commun. 326:724.
Holscher, C. (2004) Med. Microbiol. Immunol. (Berl).193:1.

Long Name

Interleukin 27

Entrez Gene IDs

246778 (Human); 246779 (Mouse); 102121242 (Cynomolgus Monkey)

Alternate Names

IL-27 p28 subunit; IL-27 subunit alpha; IL27; IL-27; IL27A; IL-27-A; IL27-A; IL27p28; IL30; interleukin 27; interleukin 30; interleukin-27 subunit alpha; MGC71873; p28IL-27A

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Citation for Human IL-27 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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