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    products/primary-antibodies/il-12a-antibody-ep5737-ab131039.pdf

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Immunology Innate Immunity Cytokines Interleukins
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RecombinantRabMAb

Recombinant Anti-IL-12A antibody [EP5737] (ab131039)

  • Datasheet
  • SDS
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
  • Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)
  • Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)
  • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
  • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
  • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
  • Anti-IL-12A antibody [EP5737] (ab131039)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP5737] to IL-12A
  • Suitable for: WB, IHC-P, Flow Cyt (Intra)
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-IL-12A antibody [EP5737]
    See all IL-12A primary antibodies
  • Description

    Rabbit monoclonal [EP5737] to IL-12A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • NIH3T3, HuT-78, and Raw 264.7 cell lysates; Human kidney tissue, Human gastric carcinoma, Human colon tissue.
  • General notes

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP5737
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-IL-12A antibody [EP5737] - BSA and Azide free (ab214433)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • NIH/3T3 whole cell lysate (ab7179)
    • RAW 264.7 whole cell lysate (ab7187)
  • Recombinant Protein

    • Recombinant Human IL-12A protein (Fc Chimera) (ab221208)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab131039 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
WB
1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P (1)
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

Flow Cyt (Intra)
1/19000.
Notes
WB
1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P
1/800. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.

Flow Cyt (Intra)
1/19000.

Target

  • Function

    Cytokine that can act as a growth factor for activated T and NK cells, enhance the lytic activity of NK/lymphokine-activated Killer cells, and stimulate the production of IFN-gamma by resting PBMC.
  • Sequence similarities

    Belongs to the IL-6 superfamily.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession P29459 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3592 Human
    • Entrez Gene: 16159 Mouse
    • Omim: 161560 Human
    • SwissProt: P29459 Human
    • SwissProt: P43431 Mouse
    • Unigene: 673 Human
    • Unigene: 103783 Mouse
    • Alternative names

      • CLMF antibody
      • CLMF p35 antibody
      • Cytotoxic lymphocyte maturation factor 1 p35 antibody
      • Cytotoxic lymphocyte maturation factor 35 kDa subunit antibody
      • IL 12 subunit p35 antibody
      • IL 12A antibody
      • IL-12 subunit p35 antibody
      • IL-12A antibody
      • IL12A antibody
      • IL12A_HUMAN antibody
      • IL35 antibody
      • IL35 subunit antibody
      • Interleukin 12 alpha chain antibody
      • Interleukin 12 p35 antibody
      • Interleukin 12 subunit alpha antibody
      • Interleukin 12A (natural killer cell stimulatory factor 1, cytotoxic lymphocyte maturation factor 1, p35) antibody
      • Interleukin-12 subunit alpha antibody
      • Natural killer cell stimulatory factor 1 35 kD subunit antibody
      • NF cell stimulatory factor chain 1 antibody
      • NFSK antibody
      • NK cell stimulatory factor chain 1 antibody
      • NKSF1 antibody
      • P35 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)

      Immunohistochemical staining of paraffin embedded human gastric carcinoma tissue section labelling IL-12A with purified ab131039 at dilution of 1/800. The secondary antibody used was HRP conjugated Goat Anti-Rabbit IgG H&L (ab97051) at dilution of 1/500. The sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

    • Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)
      Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)

      Intracellular Flow Cytometry analysis of Raw264.7 cells treated with 100ng/ml for LPS 24hour cells labelling IL-12A with ab131039 at 1/19000 dilution (1.02 µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue).

       

    • Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)
      Flow Cytometry (Intracellular) - Anti-IL-12A antibody [EP5737] (ab131039)

      Intracellular Flow Cytometry analysis of THP-1 cells treated with 100ng/ml PMA for 30min,then 1µg/ml LPS for 24hour cells labelling IL-12A with ab131039 at 1/19000 dilution (1.02µg/ml) (Red). Cells were fixed with 4% paraformaldehyde . Dylight488® (Ab98462) was used as the secondary antibody at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - Unlabelled cells (Blue). Untreated cells (Green). 

       

       

    • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      Anti-IL-12A antibody [EP5737] (ab131039) at 1/1000 dilution (purified) + HuT-78 (human Sezary syndrome) whole cell lysate at 20 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 25 kDa
      Observed band size: 35 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      All lanes : Anti-IL-12A antibody [EP5737] (ab131039) at 1/5000 dilution (purified)

      Lane 1 : NIH/3T3 (mouse embryo) whole cell lysate
      Lane 2 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

      Predicted band size: 25 kDa
      Observed band size: 35 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer 5% NFDM/TBST
    • Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      Western blot - Anti-IL-12A antibody [EP5737] (ab131039)
      All lanes : Anti-IL-12A antibody [EP5737] (ab131039) at 1/1000 dilution (unpurified)

      Lane 1 : NIH3T3 cell lysate
      Lane 2 : HuT-78 cell lysate
      Lane 3 : Raw 264.7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 25 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)

      Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labelling IL-12A with unpurified ab131039 at dilution of 1:250.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-12A antibody [EP5737] (ab131039)This image is courtesy of an anonymous Abreview.

      Formaldehyde-fixed, paraffin-embedded human colon tissue stained for IL-12A using ab131039 at 1/500 dilution in immunohistochemical analysis.

      See Abreview

    • Anti-IL-12A antibody [EP5737] (ab131039)
      Anti-IL-12A antibody [EP5737] (ab131039)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab131039? Please let us know so that we can cite the reference in this datasheet.

    ab131039 has been referenced in 7 publications.

    • Luchtel RA  et al. High-dose ascorbic acid synergizes with anti-PD1 in a lymphoma mouse model. Proc Natl Acad Sci U S A 117:1666-1677 (2020). PubMed: 31911474
    • Zhang GQ  et al. High Expression of Interleukin-12A and Its Association with the Clinicopathology and Prognosis of Differentiated Thyroid Cancer. Eur Thyroid J 9:139-147 (2020). PubMed: 32523890
    • Larousserie F  et al. Evidence for IL-35 Expression in Diffuse Large B-Cell Lymphoma and Impact on the Patient's Prognosis. Front Oncol 9:563 (2019). PubMed: 31316915
    • Liu L  et al. Periductal Mastitis: An Inflammatory Disease Related to Bacterial Infection and Consequent Immune Responses? Mediators Inflamm 2017:5309081 (2017). PubMed: 28182101
    • Zhang Y  et al. The Unknown Aspect of BAFF: Inducing IL-35 Production by a CD5+CD1dhiFc?RIIbhiRegulatory B-Cell Subset in Lupus. J Invest Dermatol 137:2532-2543 (2017). WB . PubMed: 28844943
    • Sarmento-Cabral A  et al. Metformin Reduces Prostate Tumor Growth, in a Diet-Dependent Manner, by Modulating Multiple Signaling Pathways. Mol Cancer Res 15:862-874 (2017). PubMed: 28385910
    • Terayama H  et al. Contribution of IL-12/IL-35 common subunit p35 to maintaining the testicular immune privilege. PLoS One 9:e96120 (2014). IHC-Fr ; Mouse . PubMed: 24760014

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IL-12A antibody [EP5737]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Colon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 6 for 20 mins Bond Max
    Specification
    Colon
    Blocking step
    Peroxide as per Polymer refine kit as blocking agent for 10 minute(s) · Concentration: 10% · Temperature: amb°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Feb 11 2016

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