Overview

  • Product name
    Anti-CDKN2A/p16INK4a antibody [2D9A12]
    See all CDKN2A/p16INK4a primary antibodies
  • Description
    Mouse monoclonal [2D9A12] to CDKN2A/p16INK4a
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, ELISA, IHC-Pmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human CDKN2A/p16INK4a.
    Database link: P42771

  • Positive control
    • IHC-P: Human brain tumor, ovarian carcinoma, cervix, skin and brain tissue. Rat liver tissue. Flow Cyt: HeLa cells.
  • General notes

    We have received positive and negative feedback from customers for ICC/IF. ICC/IF is not a batch-tested application for this antibody and we are not able to cover it under our Abpromise guarantee.

Properties

Applications

Our Abpromise guarantee covers the use of ab54210 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 1/50.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ELISA 1/10000.
IHC-P 1/200 - 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Recommended antigen retrieval: Place slides in a special glass slide holder and fill in the rack with slides to ensure even heating. Rinse rack in 1L of Citrate buffer in ordinary pressure cooker,(0.01M,pH6.0) to immerse all slides. Cover the cap and heat for 15-20 minutes on electric cooker until water boils. Begin timing, only 2 minutes in high-pressure Remove the pressure cooker, Allow it to cool(Cool the slides at room temperature) Rinse the slides in PBS for 5 minutes
  • Application notes
    Is unsuitable for WB.
  • Target

    • Cellular localization
      Cytoplasmic and Nuclear
    • Database links
    • Form
      There are 4 isoforms produced by alternative splicing. Isoform 1 also known as: p16INK4a; Isoform 3 also known as: p12; Isoform 4 also known as: p14ARF; p19ARF; ARF.
    • Alternative names
      • CCM2 antibody
      • CDK4 inhibitor p16 INK4 antibody
      • CDK4I antibody
      • CDKN2 antibody
      • CDKN2A antibody
      • Cell cycle negative regulator beta antibody
      • CMM2 antibody
      • Cyclin dependent kinase 4 inhibitor A antibody
      • Cyclin dependent kinase inhibitor 2A (melanoma p16 inhibits CDK4) antibody
      • Cyclin Dependent Kinase Inhibitor 2A antibody
      • Cyclin dependent kinase inhibitor 2A isoform 4 antibody
      • Cyclin dependent kinase inhibitor 2A isoforms 1/2/3 antibody
      • Cyclin dependent kinase inhibitor p16 antibody
      • INK4 antibody
      • INK4A antibody
      • MLM antibody
      • MTS1 antibody
      • Multiple tumor suppressor 1 antibody
      • p14 antibody
      • p16 antibody
      • P16INK4 antibody
      • p16INK4a antibody
      • p19 antibody
      • p19Arf antibody
      • TP16 antibody
      see all

    Images

    • Characterization of methylation and expression of p16 in the fusion cell lines.

      Confocal immunocytochemistry staining shows varying P16 expression in the various cell lines.

      Fusion cells were fixed in 4% formaldehyde/PBS for 10 minutes before a 5-minute wash in PBS. Fixed samples were blocked for 30 minutes with 0.5% Triton X-100/PBS at room temperature. Cells were incubated with ab54210 for 1 hour at 37°C, after washed with 0.5% Triton X-100/PBS, then incubated with goat-anti rabbit IgG antibody labeled with rhodamine for 1 hour at 37°C DAPI (1∶2000) was used to stain the cell nucleus.

      Photos were taken on a Leica confocal microscope.

    • ab54210 staining CDKN2A/p16INK4a in human cervix and ovarian carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

      Tissue was fixed with formaldehyde and antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/4000) for 20 minutes at 25°C. An undiluted HRP-conjugated mouse polymer was used as the secondary antibody.

      See Abreview

    • ab54210 staining CDKN2A/p16INK4a in human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

      Tissue was fixed with formaldehyde and blocked with 2% BSA for 1 hour at 22°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/100 in 1% NGS/TBS) for 20 hours at 4°C. ab6785 (1/800) was used as the secondary antibody.

      See Abreview

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in human brain tumor tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB) is shown.

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in rat liver tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB staining) is shown.

    • ab54210 at a 1:500 staining CDKN2A/p16INK4a in human brain tissue by immunohistochemistry using paraffin embedded tissue. Nuclear staining (DAB staining) is shown.

    • Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab54210 (red line).

      The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab54210, 1/50 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions.

      Acquisition of >5,000 events was performed.

      This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    References

    This product has been referenced in:
    • Kehm R  et al. Age-related oxidative changes in pancreatic islets are predominantly located in the vascular system. Redox Biol 15:387-393 (2018). Read more (PubMed: 29331666) »
    • Tsuboi T  et al. Administration of L-arginine plus L-citrulline or L-citrulline alone successfully retarded endothelial senescence. PLoS One 13:e0192252 (2018). Read more (PubMed: 29415069) »
    See all 34 Publications for this product

    Customer reviews and Q&As

    1-10 of 28 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (cartilage)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    cartilage
    Blocking step
    Bloxall then 10% Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    zinc buffered formallin

    Abcam user community

    Verified customer

    Submitted Mar 09 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (growth plate)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    growth plate
    Blocking step
    Bloxall then 10% Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    zinc buffered formallin

    Abcam user community

    Verified customer

    Submitted Mar 09 2018

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Inguinal Adipose Tissue)
    Gel Running Conditions
    Reduced Denaturing (2-20% Precast Gel, 100V for 1 hr 20 min to run)
    Loading amount
    40 µg
    Specification
    Inguinal Adipose Tissue
    Blocking step
    SuperBlock T20 (TBS) Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 23°C

    Abcam user community

    Verified customer

    Submitted Sep 13 2016

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Brain (medulloblastoma preneoplastic lesions))
    Permeabilization
    Yes - 0.3% Triton
    Specification
    Brain (medulloblastoma preneoplastic lesions)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Paraformaldehyde

    Lukas Tamayo

    Verified customer

    Submitted Jun 20 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (brain section)
    Permeabilization
    Yes - 0.8% Triton-100 in PBS
    Specification
    brain section
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Fixative
    Paraformaldehyde

    Peisu Zhang

    Verified customer

    Submitted Mar 15 2016

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (vascular smooth muscle cell)
    Permeabilization
    Yes - NP40
    Specification
    vascular smooth muscle cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Jan 06 2016

    Application
    Western blot
    Loading amount
    25 µg
    Gel Running Conditions
    Reduced Denaturing (15%gel)
    Sample
    Mouse Tissue lysate - whole (hippocampus)
    Specification
    hippocampus
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Apr 10 2015

    Answer



    CDKN2A/p16INK4a est une protéine qui est exprimé dans 63% de tumeurs et non pas que dans les tumeurs cérébrales. Vous pouvez donc également utiliser d’autres tissus cancéreux comme contrôle positif. Pour plus d’informations concernant quels tissus à utiliser comme contrôle positifs, visitez la page suivante :

    http://www.proteinatlas.org/ENSG00000147889/cancer

    Ces informations montre que la protéine est exprimé dans les cancers cervicales, les cancers de l’estomac, les cancers de la thyroïde et beaucoup plus.

    Pour plus d’informations sur la localisation de cette protéine dans des tissus normaux, regarder la publication suivante :

    Lab Invest. 1999 Sep;79(9):1137-43.
    Immunohistochemical survey of p16INK4A expression in normal human adult and infant tissues.
    Nielsen GP, Stemmer-Rachamimov AO, Shaw J, Roy JE, Koh J, Louis DN.

    Ou également, visitez le lien suivant:

    http://www.proteinatlas.org/ENSG00000147889/tissue

    Cette protéine est exprimée dans le noyau mais peut aussi être trouvé dans le cytoplasme.

    Read More

    Answer

    Thank you for contacting us.

    Human brain slides, such as ab4304, or humor brain tumor slides such as ab4679 should be good positive controls.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question
    Answer

    Thank you for contacting us.

    Here are the links to the Human Protein Atlas, including ones to the sites for normal and cancer tissue as well as cell lines. Several brain cell lines are listed as expressing CDKN2A in moderate levels.

    http://www.proteinatlas.org/ENSG00000147889
    http://www.proteinatlas.org/ENSG00000147889/normal
    http://www.proteinatlas.org/ENSG00000147889/cancer
    http://www.proteinatlas.org/ENSG00000147889/cell

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

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    1-10 of 28 Abreviews or Q&A

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