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
  • Positive control
    • Hela cells. Lymphoma This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa.

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

  • 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.

     

  • This image was kindly supplied as part of the review submitted by Joost van Galen.
  • 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:
See all 11 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Question
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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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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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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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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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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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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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 AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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