Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, ELISA Capture (Matched Antibody Pair), Neutralization

Cited:

Immunohistochemistry, Western Blot, Neutralization, Bioassay, ELISA Development, ELISA Development (Capture)

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all IL-27 Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human IL‑27 Antibody

Detection of Human IL-27 antibody by Western Blot.

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.

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

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.

Detection of Human IL-27 by Flow Cytometry

Detection of Human IL-27 by Flow Cytometry

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 (https://pubmed.ncbi.nlm.nih.gov/33584666), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human IL-27 by Flow Cytometry

Detection of Human IL-27 by Flow Cytometry

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 (https://pubmed.ncbi.nlm.nih.gov/33584666), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human IL‑27 Antibody

Application
Recommended Usage

Western Blot

1 µg/mL
Sample: MT‑2 human T cell line and CHO Chinese hamster ovary cell line transfected with human IL-27 p28

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.

Human IL-27 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human IL‑27 Biotinylated Antibody (Catalog # BAF2526)
  • Standard: Recombinant Human IL-27 Protein (Catalog # 2526-IL)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using AF2526 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

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.

Calculators

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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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

Alternate Names

IL27

Entrez Gene IDs

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

Gene Symbol

IL27

UniProt

Q14213 (EBI-3 subunit), AAM34498 (p28 subunit)

Additional IL-27 Products

Product Documents for Human IL‑27 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human IL‑27 Antibody

For research use only

Citations for Human IL‑27 Antibody

Customer Reviews for Human IL‑27 Antibody (1)

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  • Human IL-27 Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/27/2017
    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.
    Human IL‑27 Antibody AF2526

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