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RecombinantRabMAb

Recombinant Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

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Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Western blot - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Immunoprecipitation - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
  • Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP435Y-129R] to CDKN2A/p16INK4A +CDKN2B/p15INK4B - BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt (Intra), WB, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Unconjugated

Overview

  • Product name

    Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free
    See all CDKN2A/p16INK4A +CDKN2B/p15INK4B primary antibodies

  • Description

    Rabbit monoclonal [EP435Y-129R] to CDKN2A/p16INK4A +CDKN2B/p15INK4B - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK-293, HeLa ICC/IF: HeLa cells Flow Cyt (intra): HEK-293 cells. IP: HEK-293

  • General notes

    ab219723 is the carrier-free version of ab81278.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219723 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
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Predicted molecular weight: 17 kDa.
IP
Use at an assay dependent concentration.
Notes
ICC/IF
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Predicted molecular weight: 17 kDa.
IP
Use at an assay dependent concentration.
Application notes
Is unsuitable for IHC-P.

Target

  • Function

    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.

  • Tissue specificity

    Widely expressed but not detected in brain or skeletal muscle. Isoform 3 is pancreas-specific.

  • Involvement in disease

    The association between cutaneous and uveal melanomas in some families suggests that mutations in CDKN2A may account for a proportion of uveal melanomas. However, CDKN2A mutations are rarely found in uveal melanoma patients.
    Melanoma, cutaneous malignant 2
    Familial atypical multiple mole melanoma-pancreatic carcinoma syndrome
    Melanoma-astrocytoma syndrome

  • Sequence similarities

    Belongs to the CDKN2 cyclin-dependent kinase inhibitor family.
    Contains 4 ANK repeats.

  • Post-translational
    modifications

    Phosphorylation seems to increase interaction with CDK4.

  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • CDK4I antibody
      • CDKN2A antibody
      • CDN2A_HUMAN antibody
      • Cyclin-dependent kinase 4 inhibitor A antibody
      • Cyclin-dependent kinase inhibitor 2A antibody
      • MTS-1 antibody
      • Multiple tumor suppressor 1 antibody
      • p16-INK4 antibody
      • p16-INK4a antibody
      • p16INK4A antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilised HeLa (human cervix adenocarcinoma epithelial cell) labelling CDKN2A/p16INK4a  with ab81278 at 1/250 dilution, followed by AlexaFluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining in HeLa is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain alpha Tubulin at 1/200 dilution (red). The Nuclear counterstain was DAPI (blue).

      Secondary antibody only control: Secondary antibody is AlexaFluor® 488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution (green).

      This data was developed using ab81278, the same antibody clone in a different buffer formulation.

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      Flow cytometric analysis of 4% paraformaldehyde fixed and 90% methanol permeabilised HEK-293 (Human embryonic kidney epithelial cell) with ab81278 at 1/20 dilution (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

      This data was developed using ab81278, the same antibody clone in a different buffer formulation.

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      Clone EP435Y-129R (ab219723) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4A +CDKN2B/p15INK4B antibody [EP435Y-129R] (Alexa Fluor® 647). Please refer to ab199819 for protocol details.

      ab199819 staining CDKN2A/p16INK4a in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199819 at 1/200 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Western blot - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Western blot - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      All lanes : Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723) at 1/10000 dilution

      Lane 1 : HEK-293 cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 1/20 dilution per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

      Predicted band size: 17 kDa


      Exposure time: 3 seconds
    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      Clone EP435Y-129R (ab219723) has been successfully conjugated by Abcam. This image was generated using Anti-CDKN2A/p16INK4a antibody [EP435Y-129R] (Alexa Fluor® 488). Please refer to ab199756 for protocol details.

      ab199756 staining CDKN2A/p16INK4a in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199756 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293 (Human embryonic kidney epithelial cell) cells labelling CDKN2A with ab219723 at 1/100 dilution (1µg)/ 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. Gated on viable cells. 

       

       

    • Immunoprecipitation - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Immunoprecipitation - Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

      CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg with ab219723 at 1:50 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab219723 1:1000 dilution (2 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

      Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
      Lane 2: ab219723 IP in HEK-293 whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219723 in HEK-293 whole cell lysate.

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

      Exposure time: 3 seconds

    • Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)
      Anti-CDKN2A/p16INK4A + CDKN2B/p15INK4B antibody [EP435Y-129R] - BSA and Azide free (ab219723)

    References (0)

    Publishing research using ab219723? Please let us know so that we can cite the reference in this datasheet.

    ab219723 has not yet been referenced specifically in any publications.

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