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RecombinantRabMAb

Recombinant Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)

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Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
  • Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
  • Anti-CDKN2A antibody [EPR24167-43] (ab270058)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24167-43] to CDKN2A/p16INK4a
  • Suitable for: ICC/IF, IP, IHC-P, WB, Flow Cyt (Intra)
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-CDKN2A/p16INK4a antibody [EPR24167-43]
    See all CDKN2A/p16INK4a primary antibodies

  • Description

    Rabbit monoclonal [EPR24167-43] to CDKN2A/p16INK4a

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293, Saos-2, His-tagged human CDKN2A recombinant protein lysates, human CDKN2A full-length recombinant protein with His-tag, human CDKN2B full-length recombinant protein with GST-tag. IHC-P: Human cervical cancer and human endometrial cancer tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa, HEK-293 cells.

  • General notes

    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.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab270058 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
ICC/IF
1/50.
IP
1/30.
IHC-P (1)
1/500.
WB
1/1000. Predicted molecular weight: 17 kDa.
Flow Cyt (Intra)
1/500.
Notes
ICC/IF
1/50.
IP
1/30.
IHC-P
1/500.
WB
1/1000. Predicted molecular weight: 17 kDa.
Flow Cyt (Intra)
1/500.

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 antibody [EPR24167-43] (ab270058)
      Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      All lanes : Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
      Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
      Lane 3 : Saos-2 (human osteosarcoma epithelial), whole cell lysate
      Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
      Lane 5 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 17 kDa
      Observed band size: 16 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      Negative control: MCF7, MDA-MB-231 (PMID: 10914724) 

      The MW observed is consistent with the literature. (PMID: 18053084)

      This blot was developed using a higher sensitivity ECL substrate.

       Exposure time: 70 seconds.

       

    • Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      All lanes : Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution

      Lane 1 : His-tagged human CDKN2A recombinant protein
      Lane 2 : GST-tagged human CDKN2B recombinant protein

      Lysates/proteins at 0.01 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 17 kDa
      Observed band size: 16 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer and concentration: 5% NFDM/TBST

      This antibody specifically recognizes CDKN2A recombinant protein, but not CDKN2B recombinant protein.

      Exposure time: 3.25 seconds.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)

      Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human cervical cancer (PMID: 9736404). The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)

      Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human endometrial cancer. The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)

      Immunohistochemical analysis of paraffin-embedded Human cervix tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in human cervix (PMID: 9736404). The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling CDKN2A/p16INK4a with ab270058 at 1/50 (9.36 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in HeLa cell line. Negative control: MCF7 (PMID: 10914724) is observed.

      ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)

      Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 ( Human breast adenocarcinoma epithelial cell, Left)/ HeLa (Human cervix adenocarcinoma epithelial cell, Right) cells labelling CDKN2A/p16INK4a with ab270058 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: MCF7 (PMID: 10914724).

    • Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)

      CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab270058 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270058 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug

      Lane 2 : ab270058 IP in HeLa whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270058 in HeLa whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 32 seconds

       

    • Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)

      CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell), whole cell lysate 10 ug with ab270058 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270058 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HEK-293 (human embryonic kidney epithelial cell), whole cell lysate 10 ug

      Lane 2: ab270058 IP in HEK-293 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270058 in HEK-293 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 32 seconds

       

    • Anti-CDKN2A antibody [EPR24167-43] (ab270058)
      Anti-CDKN2A antibody [EPR24167-43] (ab270058)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab270058? Please let us know so that we can cite the reference in this datasheet.

    ab270058 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CDKN2A/p16INK4a antibody [EPR24167-43]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Bladder carcinoma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9
    Permeabilization
    No
    Specification
    Bladder carcinoma
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
    Fixative
    10% NBF
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Vicki Shone

    Verified customer

    Submitted Jul 13 2022

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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