Astana Biomed Group, an authorized Abcam distributor in Central Asia

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, Flow Cyt, ICC/IF
  • Reacts with: Human

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

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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: 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

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 - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
    Flow Cytometry - 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22ºC. 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 - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
    Flow Cytometry - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)

    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)

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