Human IL-27 Biotinylated Antibody

(2 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 BSA as a carrier protein.
  • Label
    Biotin
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)
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
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 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF2526) followed by Streptavidin-HRP (Catalog # DY998). Specific bands were detected for the IL‑27 EBI3/IL-27B subunit at approximately 32 kDa and IL‑27p28/IL27-A 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.
  • 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.

2 Citations: Showing 1 - 2
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Sample Type
  1. A soluble form of IL-27Ralpha is a natural IL-27 antagonist.
    Authors: Dietrich C, Candon S, Ruemmele F, Devergne O
    J Immunol, 2014;192(11):5382-9.
    Species: Human
    Sample Type: Serum
    Application: ELISA detection
  2. 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 Biotinylated Control

Ctrl BAF108 19
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Staining Reagents
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Streptavidin-Phycoerythrin

Flow F0040 7
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Streptavidin-Alkaline Phosphatase

ELISA(Det) AR001 2
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NorthernLights Streptavidin NL557

Flow, IHC, ICC NL999 4
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NorthernLights Streptavidin NL637

Flow, IHC, ICC NL998 2
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NorthernLights Streptavidin NL493

Flow, IHC, ICC NL997 1
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Streptavidin-Allophycocyanin

Flow F0050 1
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Avidin-Fluorescein

Flow F0030 1
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Supplemental Cell Selection Products
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MagCellect Streptavidin Ferrofluid

MAG999B 4
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MagCellect Streptavidin Ferrofluid

MAG999 4
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Supplemental ELISA Products
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Streptavidin-HRP

ELISA DY998 44
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EvenCoat Streptavidin Coated Plates

ELISA CP004
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EvenCoat Streptavidin Coated Plates, 5 Pack

ELISA CP003 3
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Average Rating: 4 (Based on 1 review)

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ELISA Human IL-27 Biotinylated Antibody BAF2526
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 ELISA Human Anonymous 11/27/2017
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ELISA Human IL-27 Biotinylated Antibody BAF2526
ELISA: Human IL-27 Biotinylated Antibody [BAF2526]

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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