Anti-CDKN2A/p14ARF antibody (ab3642)

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Overview

  • Product name

    Anti-CDKN2A/p14ARF antibody
    See all CDKN2A/p14ARF primary antibodies

  • Description

    Rabbit polyclonal to CDKN2A/p14ARF

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IHC-Fr, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human CDKN2A/p14ARF (C terminal). The exact sequence is proprietary.

  • Positive control

    • ICC/IF: HeLa cells. IHC-P: Lymphoma tissue.

  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab3642 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
IHC-P 1/25. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Relevance

    The gene for CDK2NA generates several transcripts/proteins which differ from each other in their first exons. Three of these transcripts are generated by alternative splicing (isoform 1 a.k.a p16INK4A, isoform 2 and isoform 3 a.k.a p12), two of which are known to function as inhibitors of CDK4 kinase. One other transcript that is generated from this gene contains an alternate reading frame (ARF), with the first exon located 20kb upstream of the remainder of the gene(isoform 4 a.k.a. p14ARF, p19ARF, ARF). In spite of the structural and some functional differences, all the proteins encoded by the CDKN2A gene are involved in cell cycle G1 control.

  • Cellular localization

    Cytoplasmic and Nuclear

  • Database links

    • Alternative names

      • ARF 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 antibody
      • INK4 antibody
      • INK4a antibody
      • Melanoma p16 inhibits CDK4 antibody
      • MLM antibody
      • MTS 1 antibody
      • MTS1 antibody
      • Multiple tumor suppressor 1 antibody
      • p14 antibody
      • p16 antibody
      • p16 INK4a antibody
      • p16INK4a antibody
      • p19 antibody
      • P19ARF antibody
      • TP16 antibody
      see all

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)

      Confocal image showing nuclear staining increased after HeLa cells transfected with CDKN2A/p14ARF.

      ICC/IF image of ab3642 stained HeLa cells. The cells were 4% paraformaldehyde fixed and then incubated in 0.1% trixtonX-100 to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with primary antibody ab3642 at a dilution of 1/250. An AlexaFluor®594 goat anti-mouse secondary IgG (ab150077) was used at a 1/1000 dilution. Anti-tubulin (ab7291) and an AlexaFluor®594 goat anti-mouse IgG (ab150120) were used as counterstains, both at a dilution of 1/1000. DAPI was used to stain the cell nuclei blue.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p14ARF antibody (ab3642)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p14ARF antibody (ab3642)
      This image was kindly supplied as part of the review submitted by Joost van Galen.
    • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)
      Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)

      ICC/IF image of ab3642 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab3642 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    References

    This product has been referenced in:

    • Cui L  et al. Overexpression of CCDC69 activates p14ARF/MDM2/p53 pathway and confers cisplatin sensitivity. J Ovarian Res 12:4 (2019). Read more (PubMed: 30651135) »
    • Sargen MR  et al. Biomarker discovery analysis: Alterations in p14, p16, p53, and BAP1 expression in nevi, cutaneous melanoma, and metastatic melanoma. Pigment Cell Melanoma Res 32:474-478 (2019). Read more (PubMed: 30672662) »
    See all 15 Publications for this product

    Publishing research using ab3642? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-7 of 7 Abreviews or Q&A

    Question

    Yes, I am interested in trying ab115663. Thanks,    

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    Answer

    Thanks so much for your reply. I am sending a free of charge vial of ab115663 on the order ***. This antibody should arrive next Thursday, the 27th. Please let me know if there is anything else that we can do for you and i"ll be happy to help.

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    Question

    Thanks for your response.  To answer your questions, my samples are human neuroblastoma cell lines.  The membranes were blocked in a 5% milk solution, and the antibody was diluted in 5% BSA.  Secondary antibody was diluted 1:2500 in 5% BSA solution. I am needing an antibody specific to the C-terminus of p14ARF; if any of your other antibodies meet this requirement I'd be happy to try them. Thanks for you help,

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    Answer

    Thank you for your response and for sending the protocol information. We also have the goat polyclonal ab115663 that is specific for the C terminus- https://www.abcam.com/CDKN2A-p14ARF-antibody-ab115663.html Would you be interested in trying this antibody?

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    Question

    Dear Abcam, I recently ordered and received ab3642 as detailed below.  However, when I used the antibody for a Western Blot two days after I received it (using the recommended dilution), I got a completely dark box (attached image).  The film was exposed for only 15 seconds and I have confirmed that the secondary antibody is working.  Is there a way that I can get either a replacement antibody or a refund since this antibody is completely unusable to me? Thanks,

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    Answer

    Thank you for contacting us and letting us know about this issue. For our documentation, could you please answer a few additional questions about your protocol? 1) What kind of samples are run on the blot (species and type)? 2) What blocking solution is used (BSA or milk)? Is the antibody also diluted in the blocking solution? 3) What concentration of secondary antibody was used? I look forward to hearing from you. We do guarantee all of our products to work as stated in tested species and applications for up to 6 months after purchase, so I would be happy to issue a refund or send a replacement. We do have other CDKN2A antibodies, such as ab470- https://www.abcam.com/CDKN2A-p14ARF-antibody-ab470.html Please let me know if you have any questions and I will be happy to help.

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    Question

    Thank you for your swift reply to my previous enquiry relating to negative control tissue. Unfortunately, I do not have access to neuropathological specimens and, in my experience, brain does not stand up well to the rigours of IHC. Therefore is there an alternative type of tissue that I could use? Many thanks.

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    Answer

    I'm afraid we are not aware of other suitable negative controls other than brain. The current literature may be able to help hopefully. Sorry I couldn't help you more on this occasion,

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    Question

    Would you be so kind as to inform me what human tissue would act as a suitable negative contol for this antibody?Many thanks.

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    Answer

    Thank you for your enquiry. I would suggest as negative control tissue to use brain. I hope this will be of help.

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    Question

    The immunogen for this antibody is a peptide. Could you please specify which p14 region is used.

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    Answer

    Thank you for your enquiry. The peptide sequence of the immunogen remains proprietary. However, I can tell you that the peptide used corresponded to the C-terminal region of Human p14ARF. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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    Question

    I want to test p14ARF (ab3642) antibody in my pilot experiment. Can I get or buy a sample/trial pack of this antibody? If yes, let me know the details.

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    Answer

    We do not routinely offer free or trial sized samples for testing purposes. Our policy at Abcam is that if an antibody does not work as specified on the datasheet, we will offer a replacement or reimbursement. Should you decide to test an antibody in an application for which we do not have any information, please let us know how you get on and in return we will forward a GBP10/ USD15/ EUR15 Amazon gift voucher.

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