RecombinantRabMAb

Recombinant Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

Reviews (6)Q&A (1)References (100)
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
  • Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1473] to CDKN2A/p16INK4a - C-terminal
  • Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free HRP PE

Overview

  • Product name

    Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal
    See all CDKN2A/p16INK4a primary antibodies

  • Description

    Rabbit monoclonal [EPR1473] to CDKN2A/p16INK4a - C-terminal

  • Host species

    Rabbit

  • Specificity

    Expression levels of the CDKN2A/p16INK4a protein may vary with sample type. It’s barely expressed in normal tissue, and mostly expressed in some tumour tissues, such as cervical cancer, breast cancer and so on. Moreover, only expressed in some cell lines. Please see images for recommended positive controls.

  • Tested applications

    Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293, HEK-293T, and Saos-2 cell lysates. IHC-P: Human cervical carcinoma tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HEK-293 and HeLa cells. IP: HeLa cell lysate.

  • General notes

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108349 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
WB (3)
1/2000. Predicted molecular weight: 17 kDa.

For unpurified use at 1/1000 - 1/10000.
IP
1/30.

For unpurified use at 1/10 - 1/100.
IHC-P (2)
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.
ICC/IF
1/270.

For unpurified use at 1/100 - 1/250.
Flow Cyt (Intra)
1/270 - 1/500.

For unpurified use at 1/100 - 1/500.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
WB
1/2000. Predicted molecular weight: 17 kDa.

For unpurified use at 1/1000 - 1/10000.
IP
1/30.

For unpurified use at 1/10 - 1/100.
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/250 - 1/500.
ICC/IF
1/270.

For unpurified use at 1/100 - 1/250.
Flow Cyt (Intra)
1/270 - 1/500.

For unpurified use at 1/100 - 1/500.  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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

    • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      All lanes : Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/1000 dilution (unpurified)

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
      Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney) cell lysate
      Lane 3 : Saos-2 (Human osteosarcoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 17 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)Image from Shan M et al., PLoS One. 2013;8(10):e76408. Fig 1.; doi: 10.1371/journal.pone.0076408. eCollection 2013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Formalin-fixed, paraffin-embedded human normal breast, luminal-A DCIS (ductal carcinoma in situ) and triple negative breast cancer tissues stained for CDKN2A/p16INK4a using ab108349 in immunohistochemical analysis.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling CDKN2A/p16INK4a with purified ab108349 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

      Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling CDKN2A/p16INK4a with unpurified ab108349 at a dilution of 1/100.

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      Overlay histogram showing HEK-293 (Human epithelial cell line from embryonic kidney) cells stained with unpurified ab108349 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab108349, 1/100) for 30 minutes at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK-293 cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions. 

       

       

    • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      All lanes : Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/2000 dilution (purified)

      Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
      Lane 2 : Saos-2 (Human osteosarcoma cell line) cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 17 kDa
      Observed band size: 17 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/2000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 17 kDa
      Observed band size: 17 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling CDKN2A/p16INK4a with unpurified ab108349 at a dilution of 1/250.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      ab108349 (purified) at 1/30 immunoprecipitating CDKN2A/p16INK4a in HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking/Dilution buffer: 5% NFDM/TBST.

       

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

      Intracellular Flow Cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling CDKN2A/p16INK4a with purified ab108349 at 1/270 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies. 

       

       

    • Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
      Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

    References (100)

    Publishing research using ab108349? Please let us know so that we can cite the reference in this datasheet.

    ab108349 has been referenced in 100 publications.

    • Si HB  et al. miR-140 Attenuates the Progression of Early-Stage Osteoarthritis by Retarding Chondrocyte Senescence. Mol Ther Nucleic Acids 19:15-30 (2020). PubMed: 31790972
    • Liu X  et al. TET2 is involved in DNA hydroxymethylation, cell proliferation and inflammatory response in keratinocytes. Mol Med Rep 21:1941-1949 (2020). PubMed: 32319620
    • Mendaza S  et al. Absence of Nuclear p16 Is a Diagnostic and Independent Prognostic Biomarker in Squamous Cell Carcinoma of the Cervix. Int J Mol Sci 21:N/A (2020). PubMed: 32204550
    • Tang H  et al. CDK5 inhibition in vitro and in vivo induces cell death in myeloma and overcomes the obstacle of bortezomib resistance. Int J Mol Med 45:1661-1672 (2020). PubMed: 32236619
    • Dungan CM  et al. In vivo analysis of ?H2AX+ cells in skeletal muscle from aged and obese humans. FASEB J 34:7018-7035 (2020). PubMed: 32246795
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-7 of 7 Abreviews or Q&A

    Western blot abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal

     Inconclusive
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Renal tubular epithelial cell)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (10%)
    Loading amount
    18 µg
    Treatment
    2 uM cisplatin 7h/d, 4day
    Specification
    Renal tubular epithelial cell
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Siyao Li

    Verified customer

    Submitted Sep 30 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (growth plate)
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: hyaluronidase
    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: 2.5% · Temperature: 23°C
    Fixative
    zinc buffered formallin
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 12 2018

    Western blot abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (vascular smooth muscle celss)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    30 µg
    Specification
    vascular smooth muscle celss
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 21 2017

    Western blot abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (Vascular smooth muscle cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    30 µg
    Specification
    Vascular smooth muscle cells
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 04 2017

    ELISA abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal

     Average
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Human Recombinant protein (Purified recombinant p16 (overexpressed in HEK293))
    Specification
    Purified recombinant p16 (overexpressed in HEK293)
    Blocking step
    SuperBlock PBS Pierce #37580 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10µg/mL · Temperature: 20°C
    Type
    Sandwich (Detection)
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 28 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CDKN2A/p16INK4a antibody [EPR1473]

     Average
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (B16F10 melanoma cancer implanted)
    Specification
    B16F10 melanoma cancer implanted
    Fixative
    4% Formalin
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: autoclaved for 10min at 121 Celcius (10mM Sodium Citrate, pH 6.0)
    Permeabilization
    Yes - 0.1% SDS
    Blocking step
    BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C
    Read More

    Ms. Seontae Kim

    Verified customer

    Submitted Mar 15 2013

    Question

    I have a customer wishing to take part in your Abtrial program.

    The information we have is as follows:

    Product Cat #: ab108349

    Application: IHC-P

    Species: Human

    Sample type: Initially in oropharyngeal squamous cell carcinoma (SCC) tissue then later in anal SCC tissue

    Read More
    Answer

    Thank you for contacting us.

    We do not provide Abtrial code for tested species and applications. Please check and explain this to customer;

    https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

    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

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com