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    products/primary-antibodies/il-27-a-antibody-epr18247-86-ab252216.pdf

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Immunology Innate Immunity Cytokines Interleukins
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RecombinantRabMAb

Recombinant Anti-IL-27-A antibody [EPR18247-86] (ab252216)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Immunoprecipitation - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
  • Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
  • Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
  • Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
  • Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
  • Anti-IL-27-A antibody [EPR18247-86] (ab252216)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18247-86] to IL-27-A
  • Suitable for: Flow Cyt (Intra), WB, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Protein
Recombinant mouse IL-27-A protein (Animal Free) (ab187220)
Primary
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Anti-DEPDC1 antibody (ab197246)

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Overview

  • Product name

    Anti-IL-27-A antibody [EPR18247-86]
    See all IL-27-A primary antibodies
  • Description

    Rabbit monoclonal [EPR18247-86] to IL-27-A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IPmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: RAW 264.7 treated with 100ng/ml LPS for 6h whole cell lysate; THP-1 whole cell lysate. Flow Cyt (intra): Daudi and HeLa cells. IP: HeLa whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18247-86
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins

Associated products

  • Alternative Versions

    • Anti-IL-27-A antibody [EPR18247-86] - BSA and Azide free (ab252381)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant mouse IL-27-A protein (Animal Free) (ab187220)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab252216 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt (Intra)
1/1000.
WB
1/1000. Predicted molecular weight: 27 kDa.
IP
1/50.
Notes
Flow Cyt (Intra)
1/1000.
WB
1/1000. Predicted molecular weight: 27 kDa.
IP
1/50.
Application notes
Is unsuitable for ICC/IF or IHC-P.

Target

  • Function

    Cytokine with pro- and anti-inflammatory properties, that can regulate T helper cell development, suppress T-cell proliferation, stimulate cytotoxic T cell activity, induce isotype switching in B-cells, and that has diverse effects on innate immune cells. Among its target cells are CD4 T helper cells which can differentiate in type 1 effector cells (TH1), type 2 effector cells (TH2) and IL17 producing helper T-cells (TH17). It drives rapid clonal expansion of naive but not memory CD4 T-cells. It also strongly synergizes with IL-12 to trigger interferon-gamma/IFN-gamma production of naive CD4 T-cells, binds to the cytokine receptor WSX-1/TCCR which appears to be required but not sufficient for IL-27-mediated signal transduction. IL-27 potentiate the early phase of TH1 response and suppress TH2 and TH17 differentiation. It induces the differentiation of TH1 cells via two distinct pathways, p38 MAPK/TBX21- and ICAM1/ITGAL/ERK-dependent pathways. It also induces STAT1, STAT3, STAT4 and STAT5 phosphorylation and activates TBX21/T-Bet via STAT1 with resulting IL12RB2 up-regulation, an event crucial to TH1 cell commitment. It suppresses the expression of GATA3, the inhibitor TH1 cells development. In CD8 T-cells, it activates STATs as well as GZMB. IL-27 reveals to be a potent inhibitor of TH17 cell development and of IL-17 production. Indeed IL-27 subunit p28 alone is also able to inhibit the production of IL17 by CD4 and CD8 T-cells. While IL-27 suppressed the development of proinflammatory Th17 cells via STAT1, it inhibits the development of anti-inflammatory inducible regulatory T-cells, iTreg, independently of STAT1. IL-27 has also an effect on cytokine production, it suppresses proinflammatory cytokine production such as IL2, IL4, IL5 and IL6 and activates suppressors of cytokine signaling such as SOCS1 and SOCS3. Apart from suppression of cytokine production, IL-27 also antagonizes the effects of some cytokines such as IL6 through direct effects on T cells. Another important role of IL-27 is its antitumor activity as well as its antiangiogenic activity with activation of production of antiangiogenic chemokines such as IP-10/CXCL10 and MIG/CXCL9. In vein endothelial cells, it induces IRF1/interferon regulatory factor 1 and increase the expression of MHC class II transactivator/CIITA with resulting up-regulation of major histocompatibility complex class II. IL-27 also demonstrates antiviral activity with inhibitory properties on HIV-1 replivation.
  • Tissue specificity

    Expressed in monocytes and in placenta.
  • Sequence similarities

    Belongs to the IL-6 superfamily.
  • Post-translational
    modifications

    O-glycosylated.
  • Cellular localization

    Secreted. Does not seem to be secreted without coexpression of EBI3.
  • Target information above from: UniProt accession Q8NEV9 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 246778 Human
    • Entrez Gene: 246779 Mouse
    • Entrez Gene: 365368 Rat
    • Omim: 608273 Human
    • SwissProt: Q8NEV9 Human
    • SwissProt: Q8K3I6 Mouse
    • Unigene: 528111 Human
    • Unigene: 222632 Mouse
    • Alternative names

      • EBI3 antibody
      • IL 27 antibody
      • IL 27 p28 subunit antibody
      • IL 27A antibody
      • IL 30 antibody
      • IL-27 subunit alpha antibody
      • IL-27-A antibody
      • IL27 antibody
      • IL27-A antibody
      • IL27A antibody
      • IL27A_HUMAN antibody
      • IL27p28 antibody
      • IL30 antibody
      • Interleukin 27 antibody
      • Interleukin 27 subunit alpha antibody
      • Interleukin 27, 28-KD subunit antibody
      • Interleukin 30 antibody
      • Interleukin-27 subunit alpha antibody
      • p28 antibody
      see all

    Images

    • Immunoprecipitation - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Immunoprecipitation - Anti-IL-27-A antibody [EPR18247-86] (ab252216)

      IL-27-A was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab252216 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab252216 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

      Lane 1: HeLa whole cell lysate 10 µg (Input).
      Lane 2: ab252216 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252216 in HeLa whole cell lysate.

      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 1 minute.

      The molecular weight observed is consistent with what has been described in the literature (PMID: 19620301, PMID: 12121660).

    • Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling IL-27-A with ab252216 at 1/100 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Flow Cytometry (Intracellular) - Anti-IL-27-A antibody [EPR18247-86] (ab252216)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Daudi (human Burkitt's lymphoma lymphoblast cell line) cells labeling IL-27-A with ab252216 at 1/100 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      All lanes : Anti-IL-27-A antibody [EPR18247-86] (ab252216) at 1/1000 dilution

      Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : Daudi (human burkitt's lymphoma lymphoblast) whole cell lysate
      Lane 3 : THP-1 (human monocytic leukemia cell line) whole cell lysate
      Lane 4 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
      Lane 5 : Rat spleen lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 27 kDa
      Observed band size: 27,48 kDa why is the actual band size different from the predicted?



      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 12121660, PMID: 21385171).

      Blocking/Dilution buffer: 5% NFDM/TBST.

      This blot was developed using a higher sensitivity ECL substrate.

      Exposure times: Lanes 1-3, 5: 3 minutes; Lane 4: 60 seconds.

    • Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Western blot - Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      All lanes : Anti-IL-27-A antibody [EPR18247-86] (ab252216) at 1/1000 dilution

      Lane 1 : Untreated RAW 264.7(mouse Abelson murine leukemia virus-induced tumor macrophage cell line) whole cell lysate
      Lane 2 : RAW 264.7 treated with 100ng/ml LPS for 6h whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 27 kDa
      Observed band size: 27,48 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 12121660, PMID: 21385171)

      Blocking/Dilution buffer: 5% NFDM/TBST.

      This blot was developed using a higher sensitivity ECL substrate.

    • Anti-IL-27-A antibody [EPR18247-86] (ab252216)
      Anti-IL-27-A antibody [EPR18247-86] (ab252216)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (1)

    Publishing research using ab252216? Please let us know so that we can cite the reference in this datasheet.

    ab252216 has been referenced in 1 publication.

    • Li Y  et al. The interplay between HIF-1a and long noncoding GAS5 regulates the JAK1/STAT3 signalling pathway in hypoxia-induced injury in myocardial cells. Cardiovasc Diagn Ther 11:422-434 (2021). PubMed: 33968620

    Customer reviews and Q&As

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