Overview

  • Product name

    Human CDKN2A ELISA Kit
    See all CDKN2A kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HEK293T Ext 5 3.6%
    Inter-assay
    Sample n Mean SD CV%
    HEK293T Ext 3 3.4%
  • Sample type

    Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.3 ng/ml
  • Range

    2.3 ng/ml - 150 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 116 110% - 125%
    Tissue Extracts 95 92% - 96%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

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

    CDKN2A in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of CDKN2A protein in human cell and tissue extracts.


    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.


    Cyclin-dependent kinase inhibitor 2A, or CDKN2A, functions as a negative regulator of the proliferation of normal cells by interacting strongly with CDK4 and CDK6. This interaction inhibits ability of CDK4 and CDK6 to interact with cyclins D and to phosphorylate the retinoblastoma protein.

  • 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
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    10X Human CDKN2A Capture Antibody 1 x 600µl
    10X Human CDKN2A Detector Antibody 1 x 600µl
    Human CDKN2A Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Relevance

    Acts as a negative regulator of the proliferation of normal cells by interacting strongly with CDK4 and CDK6. This inhibits their ability to interact with cyclins D and to phosphorylate the retinoblastoma protein. Heterodimer with CDK4 or CDK6. Predominant p16 complexes contained CDK6. Interacts (isoforms 1,2 and 4) with CDK4 (both 'T-172'-phosphorylated and non-phosphorylated forms); the interaction inhibits cyclin D-CDK4 kinase activity. Interacts with ISCO2. Widely expressed but not detected in brain or skeletal muscle. Isoform 3 is pancreas-specific.
  • Cellular localization

    Cytoplasmic and Nuclear
  • Alternative names

    • Alternative reading frame
    • ARF
    • CDK4 inhibitor p16 INK4
    • CDK4I
    • CDKN2
    • cell cycle negative regulator beta
    • CMM2
    • cyclin dependent kinase 4 inhibitor A
    • cyclin dependent kinase inhibitor 2A
    • cyclin dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
    • INK4
    • INK4A
    • MLM
    • MTS 1
    • MTS1
    • multiple tumor suppressor 1
    • P14
    • P14ARF
    • P16
    • P16 INK4A
    • P16INK4
    • P16INK4A
    • P19
    • P19ARF
    • TP16
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab227903 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 concentrations of CDKN2A were measured in duplicate and interpolated from the CDKN2A standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CDKN2A concentration was determined to be 73 ng/mL in HeLa cell extract.

  • The concentrations of CDKN2A were measured in duplicate and interpolated from the CDKN2A standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

Protocols

References

ab227903 has not yet been referenced specifically in any publications.

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