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    cdkn2ap14arf-antibody-ab3642.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors Ink4
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Anti-CDKN2A/p14ARF antibody (ab3642)

  • Datasheet
  • SDS
Submit a review Q&A (7)References (15)

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Abpromise

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Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p14ARF antibody (ab3642)
  • Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody (ab3642)

Key features and details

  • Rabbit polyclonal to CDKN2A/p14ARF
  • Suitable for: IHC-P, ICC/IF
  • Reacts with: Human
  • Isotype: IgG

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

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

    Rabbit polyclonal to CDKN2A/p14ARF
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    See all applications and species data
  • 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Ink4
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Cycle Inhibitors
    • Ink4
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • Ink4
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
Application Abreviews Notes
IHC-P
1/25. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.
Notes
IHC-P
1/25. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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

    • Entrez Gene: 1029 Human
    • Omim: 600160 Human
    • SwissProt: P42771 Human
    • SwissProt: Q8N726 Human
    • Unigene: 512599 Human
    • 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.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (15)

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

    ab3642 has been referenced in 15 publications.

    • Cui L  et al. Overexpression of CCDC69 activates p14ARF/MDM2/p53 pathway and confers cisplatin sensitivity. J Ovarian Res 12:4 (2019). 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). PubMed: 30672662
    • Liang J  et al. CDKN2A inhibits formation of homotypic cell-in-cell structures. Oncogenesis 7:50 (2018). PubMed: 29904067
    • Xie H & Wang H PRL-3 promotes breast cancer progression by downregulating p14ARF-mediated p53 expression. Oncol Lett 15:2795-2800 (2018). PubMed: 29435006
    • Yosef R  et al. Directed elimination of senescent cells by inhibition of BCL-W and BCL-XL. Nat Commun 7:11190 (2016). PubMed: 27048913
    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

    Question

    Yes, I am interested in trying ab115663. Thanks,    

    Read More

    Abcam community

    Verified customer

    Asked on Oct 20 2011

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 20 2011

    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,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 18 2011

    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?

    Read More

    Abcam Scientific Support

    Answered on Oct 18 2011

    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,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 17 2011

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 17 2011

    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.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 06 2006

    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,

    Read More

    Abcam Scientific Support

    Answered on Jun 06 2006

    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.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 04 2006

    Answer

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

    Read More

    Abcam Scientific Support

    Answered on Jun 06 2006

    Question

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

    Read More

    Abcam community

    Verified customer

    Asked on Dec 23 2005

    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.

    Read More

    Abcam Scientific Support

    Answered on Dec 23 2005

    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.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 13 2003

    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.

    Read More

    Asdf Edo

    Abcam Scientific Support

    Answered on Oct 13 2003

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

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