Human IL-27 Antibody

(7 citations)
(1 Review)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-27 in ELISAs and Western blots. In sandwich immunoassays, less than 0.2% cross-reactivity with recombinant mouse IL‑27, recombinant human (rh) IL‑12, and rhIL-12 p40 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human IL‑27 (R&D Systems, Catalog # 2526-IL)
    Arg21-Lys229 (IL-27 EBI-3 subunit), Phe29-Pro243 (IL-27 p28 subunit)
    Accession # Q14213 (EBI-3 subunit), AAM34498 (p28 subunit)
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
    • Human IL-27 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human IL‑27 Antibody (Catalog # AF2526)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human IL‑27 Biotinylated Antibody (Catalog # BAF2526)
  • ELISA Standard
     
    Recombinant Human IL-27 Protein (Catalog # 2526-IL)
  • Neutralization
    Measured by its ability to neutralize IL‑27 inhibition of EMCV-induced cytopathy in the HepG2 human hepatocellular carcinoma cell line. Bender, H. et al. (2009) Hepatology 50:585. The Neutralization Dose (ND50) is typically 1‑5 µg/mL in the presence of 25 ng/mL Recombinant Human IL‑27.
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.
Data Examples
IL‑27 Inhibition of EMCV-induced Cytopathy and Neutralization by Human IL‑27 Antibody. Recombinant Human IL‑27 (Catalog # 2526‑IL) reduces the Encephalo­myocarditis Virus (EMCV)-induced cytopathy in the HepG2 human hepatocellular carcinoma cell line in a dose-dependent manner (orange line). Inhibition of EMCV activity elicited by Recombinant Human IL‑27 (25 ng/mL) is neutralized (green line) by increasing concen­trations of Goat Anti-Human IL‑27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2526). The ND50 is typically 1‑5 µg/mL.
Detection of Human IL‑27 by Western Blot. Western blot shows lysates of MT‑2 human T cell line and CHO Chinese hamster ovary cell line either mock transfected or transfected with human IL-27 p28/IL-27A. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human IL‑27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2526) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for the IL‑27 EBI3/IL‑27B at approximately 32 kDa and IL‑27 p28/IL-27A at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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), also known as IL27B, a 34 kDa glycoprotein that is related to the p40 subunit of IL-12 and IL-23, and p28, also known as IL27A, 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:
    1. Boulay, J-L. et al. (2003) Immunity 19:159. 
    2. Trinchieri, G. et al. (2003) Immunity 19:641. 
    3. Murakami, M. et al. (2004) Growth Factors 22:75. 
    4. Cordoba-Rodriguez, R. and D.M. Frucht (2003) Exp. Opin. Biol. Ther. 3:715. 
    5. Devergne, O. et al. (1996) J. Virology 70:1143. 
    6. Pflanz, S. et al. (2002) Immunity 16:779.
    7. Villarino, A.V. et al. (2004) J. Immunol. 173:715.
    8. Pflanz, S. et al. (2004) J Immunol 172:2225.
    9. Scheller, J. et al. (2005) Biochem. Biophys. Res. Commun. 326:724.
    10. Holscher, C. (2004) Med. Microbiol. Immunol. (Berl).193:1.
  • Long Name:
    Interleukin 27
  • Entrez Gene IDs:
    246778 (Human); 246779 (Mouse)
  • 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
Related Research Areas
Citations:

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.

7 Citations: Showing 1 - 7
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Species
Applications
Sample Type
  1. Type I IFN augments IL-27-dependent TRIM25 expression to inhibit HBV replication
    Authors: G Tan, Q Xiao, H Song, F Ma, F Xu, D Peng, N Li, X Wang, J Niu, P Gao, FX Qin, G Cheng
    Cell. Mol. Immunol, 2017;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4
    Authors: A Garg, R Trout, SA Spector
    Sci Rep, 2017;7(0):44485.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. HIV and HIV-Tat inhibit LPS-induced IL-27 production in human macrophages by distinct intracellular signaling pathways
    Authors: N Gajanayaka, S O'Hara, Y Konarski, J Fernandes, K Muthumani, M Kozlowski, JB Angel, A Kumar
    J. Leukoc. Biol., 2017;0(0):.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Capture
  4. IL-27 and type 2 immunity in asthmatic patients: association with severity, CXCL9, and signal transducer and activator of transcription signaling.
    Authors: Xie M, Mustovich A, Jiang Y, Trudeau J, Ray A, Ray P, Hu H, Holguin F, Freeman B, Wenzel S
    J Allergy Clin Immunol, 2015;135(2):386-94.
    Species: Human
    Sample Type: Whole Cells
    Application: IHC Not Specified
  5. Independent and interdependent immunoregulatory effects of IL-27, IFN-beta, and IL-10 in the suppression of human Th17 cells and murine experimental autoimmune encephalomyelitis.
    Authors: Fitzgerald D, Fonseca-Kelly Z, Cullimore M, Safabakhsh P, Saris C, Zhang G, Rostami A
    J Immunol, 2013;190(7):3225-34.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  6. B cells as a therapeutic target for IFN-beta in relapsing-remitting multiple sclerosis.
    Authors: Ramgolam VS, Sha Y, Marcus KL, Choudhary N, Troiani L, Chopra M, Markovic-Plese S
    J. Immunol., 2011;186(0):4518.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  7. Suppression of TNF-alpha and IL-1 signaling identifies a mechanism of homeostatic regulation of macrophages by IL-27.
    Authors: Kalliolias GD, Gordon RA, Ivashkiv LB
    J. Immunol., 2010;185(11):7047-56.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Development
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
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Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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Average Rating: 4 (Based on 1 review)

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ELISA Human IL-27 Antibody AF2526
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 ELISA Human Anonymous 11/27/2017
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ELISA Human IL-27 Antibody AF2526
ELISA: Human IL-27 Antibody [AF2526]

Summary

ApplicationELISA
Sample TestedSerum and Plasma
SpeciesHuman

Other Experimental Details

Other Experimental DetailsAF2526 was used as the solid phase antibody, BAF2526 was used as the detection antibody. 2526-IL was used as the immunoassay standard. Sensitivity of the assay was around 100 pg/ml.

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