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Human IL-27 Antibody

Catalog #: AF2526
13 Citations 1 Review Datasheet / COA / SDS
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AF2526
AF2526-SP
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Detection of Human IL‑27 by Western Blot.
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Citations (13)
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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 ELISAs and Western blots.
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 either lyophilized or 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

Recommended Concentration
Reagent
ELISA Capture (Matched Antibody Pair)
0.2-0.8 µg/mL 

Use in combination with:

Detection Reagent: Human IL‑27 Biotinylated Antibody (Catalog # BAF2526)

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 0.1‑0.8 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.

Scientific Data

Western Blot Detection of Human IL-27 antibody by Western Blot. View Larger

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 (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.

Neutralization IL‑27 Inhibition of EMCV-induced Cytopathy and Neutralization by Human IL‑27 Antibody. View Larger

IL‑27 Inhibition of EMCV-induced Cytopathy and Neutralization by Human IL‑27 Antibody. Recombinant Human IL-27 (Catalog # 2526-IL) reduces the Encephalo-myocarditis 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 concen-trations of Goat Anti-Human IL-27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2526). The ND50 is typically 0.1-0.8 µg/mL.

Reconstitution Calculator

Reconstitution Calculator

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.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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); 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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Citations 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.

13 Citations: Showing 1 - 10
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  1. Human interleukin-12? and EBI3 are cytokines with anti-inflammatory functions
    Authors: Hildenbrand, K;Bohnacker, S;Menon, PR;Kerle, A;Prodjinotho, UF;Hartung, F;Strasser, PC;Catici, DAM;Rührnö beta l, F;Haslbeck, M;Schumann, K;Müller, SI;da Costa, CP;Esser-von Bieren, J;Feige, MJ;
    Science advances
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  2. Fungal sensing by dectin-1 directs the non-pathogenic polarization of TH17 cells through balanced type I IFN responses in human DCs
    Authors: SI Gringhuis, TM Kaptein, EBM Remmerswaa, A Drewniak, BA Wevers, B Theelen, GRAM D'Haens, T Boekhout, TBH Geijtenbee
    Nature Immunology, 2022-12-01;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. IL-27 promotes decidualization via the STAT3-ESR/PGR regulatory axis
    Authors: XY Zhang, HH Shen, XY Qin, CJ Wang, WT Hu, SP Liu, JN Wu, F Xie, FY Xu, SM Zhao, YY Yuan, MQ Li
    Journal of reproductive immunology, 2022-04-12;151(0):103623.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  4. RIG-I-like receptor activation by dengue virus drives follicular T helper cell formation and antibody production
    Authors: JK Sprokholt, TM Kaptein, JL van Hamme, RJ Overmars, SI Gringhuis, TBH Geijtenbee
    PLoS Pathog., 2017-11-29;13(11):e1006738.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Interleukin 27 is increased in carotid atherosclerosis and promotes NLRP3 inflammasome activation
    Authors: I Gregersen, Ø Sandanger, ET Askevold, EL Sagen, K Yang, S Holm, TM Pedersen, M Skjelland, K Krohg-Søre, TV Hansen, TB Dahl, K Otterdal, T Espevik, P Aukrust, A Yndestad, B Halvorsen
    PLoS ONE, 2017-11-27;12(11):e0188387.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  6. 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., 2017-07-11;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development (Capture)
  7. 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, 2017-03-24;7(0):44485.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  8. 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, 2017-02-13;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  9. 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, 2014-10-11;135(2):386-94.
    Species: Human
    Sample Types: Whole Cells
    Applications: IHC
  10. Fucose-based PAMPs prime dendritic cells for follicular T helper cell polarization via DC-SIGN-dependent IL-27 production.
    Authors: Gringhuis S, Kaptein T, Wevers B, van der Vlist M, Klaver E, van Die I, Vriend L, de Jong M, Geijtenbeek T
    Nat Commun, 2014-10-03;5(0):5074.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  11. 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, 2013-03-01;190(7):3225-34.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  12. 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., 2011-03-02;186(0):4518.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  13. 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., 2010-10-22;185(11):7047-56.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development

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Human IL-27 Antibody
By Anonymous on 11/27/2017
Application: ELISA Sample Tested: Serum and Plasma Species: Human

AF2526 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.

ELISA Human IL-27 Antibody AF2526