IL-27 Antibody [Unconjugated]

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.

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.

IL-27 secreted by TFH cells support plasmablasts and plasma cell formation. (A–C) Mechanism of TFH mediated B cell response was determined by autologous TFH-B cell co-culture (CHB: n=5, HC-vacc: n=5). To imitate HBV-specific interactions between TFH and B cells, FACS-sorted TFH cells were first primed with HBsAg for 3 h, washed and then incubated with autologous CD19+CD27+ memory and CD19+CD27-IgD+ naïve B cells with and without of IL-21 and IL-27 neutralizing antibodies for 5 days. Generation of plasmablasts and plasma cells was analyzed by flow cytometry (D) expression of Blimp-1 (CHB: n=5, HC-vacc: n=5). Plasmablasts were gated as CD27+CD38+ cells and plasma cells were defined based on CD27+CD38+CD138. (E) Incubation of memory B and naïve B cells with rIL-27 for 5 days showed increased plasmablasts and plasma cell formation (CHB: n=5). Statistical analysis was performed using either Kruskal-Wallis test (ANOVA) with Dunn’s post hoc test for multiple comparisons or paired t test. Bars indicates mean and error bars designate standard deviation. * indicates p < 0.05, **p < 0.01 and ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33584666), licensed under a CC-BY license. Not internally tested by R&D Systems.

IL-27 secreted by TFH cells support plasmablasts and plasma cell formation. (A–C) Mechanism of TFH mediated B cell response was determined by autologous TFH-B cell co-culture (CHB: n=5, HC-vacc: n=5). To imitate HBV-specific interactions between TFH and B cells, FACS-sorted TFH cells were first primed with HBsAg for 3 h, washed and then incubated with autologous CD19+CD27+ memory and CD19+CD27-IgD+ naïve B cells with and without of IL-21 and IL-27 neutralizing antibodies for 5 days. Generation of plasmablasts and plasma cells was analyzed by flow cytometry (D) expression of Blimp-1 (CHB: n=5, HC-vacc: n=5). Plasmablasts were gated as CD27+CD38+ cells and plasma cells were defined based on CD27+CD38+CD138. (E) Incubation of memory B and naïve B cells with rIL-27 for 5 days showed increased plasmablasts and plasma cell formation (CHB: n=5). Statistical analysis was performed using either Kruskal-Wallis test (ANOVA) with Dunn’s post hoc test for multiple comparisons or paired t test. Bars indicates mean and error bars designate standard deviation. * indicates p < 0.05, **p < 0.01 and ***p < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33584666), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, B/N, ELISA(Cap)
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

IL-27 Antibody [Unconjugated] Summary

• 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)
• Specificity: Detects human IL-27 in ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: IL27
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Neutralization
  • Western Blot 1 ug/mL
• Application Notes: ELISA Capture: Human IL-27 Antibody (Catalog # AF2526)
  ELISA Detection: Human IL-27 Biotinylated Antibody (Catalog # BAF2526)
  Standard: Recombinant Human IL-27 (Catalog # 2526-IL)

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-27 Antibody [Unconjugated]

• 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

Background

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).

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-27 Antibody (AF2526)(18)

Publications using AF2526

• 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; Human interleukin-12? and EBI3 are cytokines with anti-inflammatory functions Science advances 2023-10-27 [PMID: 37878703] (Western Blot, Human)
• SI Gringhuis, TM Kaptein, EBM Remmerswaa, A Drewniak, BA Wevers, B Theelen, GRAM D'Haens, T Boekhout, TBH Geijtenbee Fungal sensing by dectin-1 directs the non-pathogenic polarization of TH17 cells through balanced type I IFN responses in human DCs Nature Immunology, 2022-12-01;0(0):. 2022-12-01 [PMID: 36456734] (Neutralization, Human)
• XY Zhang, HH Shen, XY Qin, CJ Wang, WT Hu, SP Liu, JN Wu, F Xie, FY Xu, SM Zhao, YY Yuan, MQ Li IL-27 promotes decidualization via the STAT3-ESR/PGR regulatory axis Journal of reproductive immunology, 2022-04-12;151(0):103623. 2022-04-12 [PMID: 35430461] (Neutralization, Human)
• Yingxia Zheng, Li Han, Zheyi Chen, Yiyang Li, Bingqian Zhou, Rui Hu et al. PD-L1+CD8+ T cells enrichment in lung cancer exerted regulatory function and tumor-promoting tolerance iScience 2022-02-18 [PMID: 35146396]
• Lei V, Handfield C, Kwock JT et al. Skin Injury Activates A Rapid TRPV1-dependent Antiviral Protein Response The Journal of investigative dermatology 1/7/2022 [PMID: 35007556]
• Richard A O’Connor, Vishwani Chauhan, Layla Mathieson, Helen Titmarsh, Lilian Koppensteiner, Irene Young et al. T cells drive negative feedback mechanisms in cancer associated fibroblasts, promoting expression of co-inhibitory ligands, CD73 and IL-27 in non-small cell lung cancer OncoImmunology 2021-07-22 [PMID: 34290905]
• Arshi Khanam, Natarajan Ayithan, Lydia Tang, Bhawna Poonia, Shyam Kottilil IL-21–Deficient T Follicular Helper Cells Support B Cell Responses Through IL-27 in Patients With Chronic Hepatitis B Frontiers in Immunology 2021-01-28 [PMID: 33584666]
• JK Sprokholt, TM Kaptein, JL van Hamme, RJ Overmars, SI Gringhuis, TBH Geijtenbee RIG-I-like receptor activation by dengue virus drives follicular T helper cell formation and antibody production PLoS Pathog., 2017-11-29;13(11):e1006738. 2017-11-29 [PMID: 29186193] (Neutralization, Human)
• 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 Interleukin 27 is increased in carotid atherosclerosis and promotes NLRP3 inflammasome activation PLoS ONE, 2017-11-27;12(11):e0188387. 2017-11-27 [PMID: 29176764] (Neutralization, Human)
• N Gajanayaka, S O'Hara, Y Konarski, J Fernandes, K Muthumani, M Kozlowski, JB Angel, A Kumar HIV and HIV-Tat inhibit LPS-induced IL-27 production in human macrophages by distinct intracellular signaling pathways J. Leukoc. Biol., 2017-07-11;0(0):. 2017-07-11 [PMID: 28698313] (ELISA Development (Capture), Human)
• A Garg, R Trout, SA Spector Human Immunodeficiency Virus Type-1 Myeloid Derived Suppressor Cells Inhibit Cytomegalovirus Inflammation through Interleukin-27 and B7-H4 Sci Rep, 2017-03-24;7(0):44485. 2017-03-24 [PMID: 28338007] (Neutralization, Human)
• G Tan, Q Xiao, H Song, F Ma, F Xu, D Peng, N Li, X Wang, J Niu, P Gao, FX Qin, G Cheng Type I IFN augments IL-27-dependent TRIM25 expression to inhibit HBV replication Cell. Mol. Immunol, 2017-02-13;0(0):. 2017-02-13 [PMID: 28194021] (Neutralization, Human)
• Xie M, Mustovich A, Jiang Y, Trudeau J, Ray A, Ray P, Hu H, Holguin F, Freeman B, Wenzel S IL-27 and type 2 immunity in asthmatic patients: association with severity, CXCL9, and signal transducer and activator of transcription signaling. J Allergy Clin Immunol, 2014-10-11;135(2):386-94. 2014-10-11 [PMID: 25312760] (IHC, Human)
• Gringhuis S, Kaptein T, Wevers B, van der Vlist M, Klaver E, van Die I, Vriend L, de Jong M, Geijtenbeek T Fucose-based PAMPs prime dendritic cells for follicular T helper cell polarization via DC-SIGN-dependent IL-27 production. Nat Commun, 2014-10-03;5(0):5074. 2014-10-03 [PMID: 25278262] (Neutralization, Human)
• Jordan V. Price, David J. Haddon, Dodge Kemmer, Guillaume Delepine, Gil Mandelbaum, Justin A. Jarrell et al. Protein microarray analysis reveals BAFF-binding autoantibodies in systemic lupus erythematosus Journal of Clinical Investigation 2013-12-02 [PMID: 24270423] (Bioassay, N/A)
• Fitzgerald D, Fonseca-Kelly Z, Cullimore M, Safabakhsh P, Saris C, Zhang G, Rostami A 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. J Immunol, 2013-03-01;190(7):3225-34. 2013-03-01 [PMID: 23455508] (Neutralization, Human)
• Ramgolam VS, Sha Y, Marcus KL, Choudhary N, Troiani L, Chopra M, Markovic-Plese S B cells as a therapeutic target for IFN-beta in relapsing-remitting multiple sclerosis. J. Immunol., 2011-03-02;186(0):4518. 2011-03-02 [PMID: 21368231] (Neutralization, Human)
• Kalliolias GD, Gordon RA, Ivashkiv LB Suppression of TNF-alpha and IL-1 signaling identifies a mechanism of homeostatic regulation of macrophages by IL-27. J. Immunol., 2010-10-22;185(11):7047-56. 2010-10-22 [PMID: 20971923] (ELISA Development, Human)

Bioinformatics

• Gene Symbol: IL27
• Uniprot:
  • Q14213 (EBI-3 subunit), AAM34498 (p28 subunit)()