Overview

  • Product name

    Human IL-12/IL-23 p40 ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Supernatant 5 3.6%
    Inter-assay
    Sample n Mean SD CV%
    Supernatant 3 2.3%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    5.8 pg/ml
  • Range

    31.3 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 101 95% - 105%
    Cell culture media 109 101% - 115%
    Heparin Plasma 107 101% - 113%
    EDTA Plasma 101 96% - 105%
    Citrate Plasma 101 99% - 103%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    IL-12/IL-23p40 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-12/IL-23P40 protein in human 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 substrate 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.


     

  • Notes

    IL-12/IL-23P40 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, as well as the heterodimerized forms.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human IL-12B/IL-23p40 Detector Antibody 1 x 600µl
    10X Human IL-12B/IL-23 p40 Capture Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent CPI 1 x 6ml
    Human IL-12B/IL-23p40 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 x 96 tests
    Sample Diluent NS 1 x 50ml
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    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.
  • Alternative names

    • CLMF
    • CLMF p40
    • CLMF2
    • Cytotoxic lymphocyte maturation factor 2
    • Cytotoxic lymphocyte maturation factor 40 kDa subunit
    • IL 12 subunit p40
    • IL 12B
    • IL12 + IL23 p40
    • IL12 subunit p40
    • IL12B
    • Interleukin 12 beta chain
    • Interleukin 12 p40
    • Interleukin 12 subunit beta
    • Interleukin 12B
    • Natural killer cell stimulatory factor 2
    • Natural killer cell stimulatory factor 40 kD subunit
    • NK cell stimulatory factor chain 2
    • NKSF
    • NKSF2
    • p40
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab220656 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.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentration of IL-12/IL-23P40 was measured in duplicate, interpolated from the IL-12/IL-23P40 standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (heparin) 50% and plasma (citrate) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of IL-12/IL-23P40 were measured in duplicate, interpolated from the IL-12/IL-23P40 standard curve and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (EDTA) 50%, plasma (heparin) 50% and plasma (citrate) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-12/IL-23P40 concentration was determined to be 45 pg/mL in serum, 41 pg/mL in plasma (EDTA), 42 pg/mL in plasma (heparin) and 42 pg/mL in plasma (citrate).

  • Serum from ten individual healthy human female donors was measured in duplicate. Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-12/IL-23P40 concentration was determined to be  92 pg/mL with a range of 17 – 100 pg/mL.

  • The concentrations of IL-12/IL-23P40 was measured in duplicate, interpolated from the IL-12/IL-23P40 standard curve and corrected for sample dilution. Undiluted samples are as follows:  Stimulated PBMC cell culture supernatant (50%) and Stimulated THP1 cell culture supernatant (12.5%).  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-12/IL-23P40 concentration was determined to be 369 pg/mL in Stimulated PBMC Cell culture supernatant and 10,253 pg/mL in THP1 cell culture supernatant.  IL-12/IL-23P40 was undetectable in media or unstimulated supernatant samples.

Protocols

References

ab220656 has not yet been referenced specifically in any publications.

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