Overview

  • Product name

    Mouse IL-12 p40 ELISA Kit
    See all IL-12 p40 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 5 3.5%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 3.1%
  • Sample type

    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich
  • Sensitivity

    3.7 pg/ml
  • Range

    15.6 pg/ml - 1000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 115 108% - 124%
    Cell culture media 94 92% - 96%
    Heparin Plasma 107 103% - 113%
    EDTA Plasma 122 118% - 125%
    Citrate Plasma 111 105% - 114%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Predicted to work with: RatDoes not react with: Human
  • Product overview

    IL-12 p40 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-12 p40 protein in mouse serum, plasma, and cell culture supernatant.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    IL-12 p40 is a 40kDa cytokine that exists in its monomeric form as well as heterodimerizes with a 35kDa subunit to form IL-12/IL-12p70 and heterodimerizes with a 19kDa subunit to form IL-23.  IL-12/IL-23P40 can act as a growth factor for activated T and natural killer cells as well as stimulate the production of Interferon gamma.  IL-12/IL-23P40 is expressed by activated macrophages that induces Th1 cell development.  This assay recognizes IL-12/IL-23P40 monomer and homodimer.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 4BR 1 x 6ml
    10X Mouse IL-12 p40 Capture Antibody 1 x 600µl
    10X Mouse IL-12 p40 Detector Antibody 1 x 600µl
    Mouse IL-12 p40 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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, an 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 an 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
  • Alternative names

    • CLMF
    • CLMF p40
    • CLMF2
    • Cytotoxic lymphocyte maturation factor 40 kDa subunit
    • IL 12 subunit p40
    • IL 12B
    • IL-12 subunit p40
    • IL-12B
    • IL12 p40
    • IL12 subunit p40
    • IL12B
    • IL12B_HUMAN
    • interleukin 12 beta chain
    • Interleukin 12 p40
    • Interleukin 12 subunit beta
    • Interleukin 12B
    • Interleukin-12 subunit beta
    • natural killer cell stimulatory factor 40 kD subunit
    • NK cell stimulatory factor chain 2
    • NKSF
    • NKSF2
    • p40
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab236717 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The IL-12 p40 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of IL-12 p40 were measured in duplicates, interpolated from the IL-12 p40 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA) 50%, and plasma (heparin) 50%.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-12 p40 concentration was determined to be 1509 pg/mL in serum, 856 pg/mL in plasma (citrate), 822 pg/mL in plasma (EDTA) and 2285 pg/mL in plasma (heparin).

  • The concentrations of IL-12 p40 were measured in duplicates, interpolated from the IL-12 p40 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Protocols

References

This product has been referenced in:

  • Zhuge X  et al. Acetate metabolic requirement of avian pathogenic Escherichia coli promotes its intracellular proliferation within macrophage. Vet Res 50:31 (2019). Read more (PubMed: 31046828) »
See 1 Publication for this product

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