Overview

  • Product name

    Anti-IL-12B antibody [EPR5739]
    See all IL-12B primary antibodies
  • Description

    Rabbit monoclonal [EPR5739] to IL-12B
  • Host species

    Rabbit
  • Specificity

    This antibody is unsuitable to detect endogenous IL-12B in WB.

  • Tested applications

    Suitable for: Flow Cyt, WB, ICCmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human IL-12B aa 300-400. The exact sequence is proprietary.
    Database link: P29460

  • Positive control

    • WB: Human IL-12B recombinant protein. ICC/IF: MOLT-4 cells Flow Cyt: Jurkat cells.
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133752 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 1/300.
WB 1/1000 - 1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).

This antibody is unsuitable to detect endogenous IL12B in WB

ICC 1/100 - 1/250.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • 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.
      Associates with IL23A to form the IL-23 interleukin, a heterodimeric cytokine which functions in innate and adaptive immunity. IL-23 may constitute with IL-17 an acute response to infection in peripheral tissues. IL-23 binds to a heterodimeric receptor complex composed of IL12RB1 and IL23R, activates the Jak-Stat signaling cascade, stimulates memory rather than naive T-cells and promotes production of proinflammatory cytokines. IL-23 induces autoimmune inflammation and thus may be responsible for autoimmune inflammatory diseases and may be important for tumorigenesis.
    • Involvement in disease

      Defects in IL12B are a cause of Mendelian susceptibility to mycobacterial disease (MSMD) [MIM:209950]; also known as familial disseminated atypical mycobacterial infection. This rare condition confers predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections, with the exception of Salmonella which infects less than 50% of these individuals. The pathogenic mechanism underlying MSMD is the impairment of interferon-gamma mediated immunity, whose severity determines the clinical outcome. Some patients die of overwhelming mycobacterial disease with lepromatous-like lesions in early childhood, whereas others develop, later in life, disseminated but curable infections with tuberculoid granulomas. MSMD is a genetically heterogeneous disease with autosomal recessive, autosomal dominant or X-linked inheritance.
      Genetic variations in IL12B are a cause of susceptibility to psoriasis type 11 (PSORS11) [MIM:612599]. Psoriasis is a common, chronic inflammatory disease of the skin with multifactorial etiology. It is characterized by red, scaly plaques usually found on the scalp, elbows and knees. These lesions are caused by abnormal keratinocyte proliferation and infiltration of inflammatory cells into the dermis and epidermis.
    • Sequence similarities

      Belongs to the type I cytokine receptor family. Type 3 subfamily.
      Contains 1 fibronectin type-III domain.
      Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    • Post-translational
      modifications

      Known to be C-mannosylated in the recombinant protein; it is not yet known for sure if the wild-type protein is also modified.
    • Cellular localization

      Secreted.
    • Information by UniProt
    • Database links

    • Alternative names

      • CLMF antibody
      • CLMF p40 antibody
      • CLMF2 antibody
      • Cytotoxic lymphocyte maturation factor 40 kDa subunit antibody
      • IL 12B antibody
      • IL-12 subunit p40 antibody
      • IL-12B antibody
      • IL12 p40 antibody
      • IL12B antibody
      • IL12B_HUMAN antibody
      • IMD28 antibody
      • IMD29 antibody
      • Interleukin 12 beta chain antibody
      • Interleukin 12 p40 antibody
      • Interleukin-12 subunit beta antibody
      • NK cell stimulatory factor chain 2 antibody
      • NKSF antibody
      • NKSF2 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence analysis of MOLT-4 (Human lymphoblastic leukemia T lymphoblast) cells labeling IL-12B with Purified ab133752 at 1:100 dilution (3.3 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were not counterstained. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • All lanes : Anti-IL-12B antibody [EPR5739] (ab133752) at 1/1000 dilution (Purified)

      All lanes : Human IL-12 recombinant protein

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 36 kDa
      Observed band size: 40 kDa
      why is the actual band size different from the predicted?



      The recombinant protein contains Human IL-12B (Ile23-Ser328) and Human IL-12A (Arg23-Ser219) connected by disulfide bond to be heterodimer. Under reducing conditions, the observed bands will be at 40 KD for IL-12B and 29 KD for IL-12A.

    • Anti-IL-12B antibody [EPR5739] (ab133752) at 1/1000 dilution + Human IL-12B recombinant protein at 0.01 µg

      Secondary
      Goat anti-Rabbit HRP at 1/2000 dilution

      Predicted band size: 36 kDa
      Observed band size: 40 kDa why is the actual band size different from the predicted?



      This image was generated using the unpurified version of the product. 

    • Flow cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling IL-12B (red) with ab133752 at a 1/2500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

       

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Reitberger S  et al. Assembly-induced folding regulates interleukin 12 biogenesis and secretion. J Biol Chem 292:8073-8081 (2017). Read more (PubMed: 28325840) »
    See 1 Publication for this product

    Customer reviews and Q&As

    Answer


    The lab let me know that they did not prepare the sample ourselves, rather we used ab9993 (https://www.abcam.com/index.html?datasheet=9993)

    We loaded 10 ng/well and used a 1:5,000 primary dilution. We used our standard SDS-PAGE gel for reducing conditions.

    Read More

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