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-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human IL-12A aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

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

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

    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.

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

Applications

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 1/800. 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.

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.
  • Information by UniProt
  • Database links

  • 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

  • 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 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 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 (1.02 µg/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)
  • 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
  • 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
  • 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

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

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

    See Abreview

References

This product has been referenced in:

  • Liu L  et al. Periductal Mastitis: An Inflammatory Disease Related to Bacterial Infection and Consequent Immune Responses? Mediators Inflamm 2017:5309081 (2017). Read more (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 . Read more (PubMed: 28844943) »
See all 4 Publications for this product

Customer reviews and Q&As

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

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Verified customer

Submitted Feb 11 2016

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