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Cell Biology Cell Cycle Cell Cycle Inhibitors Ink4

Anti-CDKN2A/p19ARF抗体(ab80)

  • Datasheet
  • SDS
Reviews (15)Q&A (24)References (212)

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Western blot - Anti-CDKN2A/p19ARF antibody (ab80)
  • Immunocytochemistry - Anti-CDKN2A/p19ARF antibody (ab80)

Key features and details

  • Rabbit polyclonal to CDKN2A/p19ARF
  • Suitable for: ICC/IF, IHC-Fr, ELISA, ICC, WB, IHC-P
  • Reacts with: Mouse
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-CDKN2A/p16INK4a antibody [EPR20418] (ab211542)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-CDKN2A/p19ARF抗体
    参阅全部 CDKN2A/p19ARF 一抗
  • 描述

    兔多克隆抗体to CDKN2A/p19ARF
  • 宿主

    Rabbit
  • 特异性

    On Western blot the antibody reveals IVT murine p19ARF and a band of the same size in CTLL2 cells. It detects a specific band in WT mouse embryo fibroblasts which is not present in p19ARF-null MEFs.
  • 经测试应用

    适用于: ICC/IF, IHC-Fr, ELISA, ICC, WB, IHC-Pmore details
  • 种属反应性

    与反应: Mouse
    不与反应: Rat, Human
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 常规说明

    A monoclonal alternative to this target is available (ab26696).

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 纯化说明

    The serum has been purified on a peptide-NHS-activated Sepharose column. Specific antibodies have been eluted with 100 mM glycine pH 2.5. The purified antibodies have been dialysed against PBS pH 7.4.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Ink4
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Epigenetics and Nuclear Signaling
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    • Cell Cycle Inhibitors
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    • Cell cycle inhibitors
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    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • INK4a/Arf

相关产品

  • 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)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab80于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF (3)
Use at an assay dependent concentration.
IHC-Fr (2)
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC
Use a concentration of 1 µg/ml. Fix with a 1:1 mixture of methanol:acetone at -20C for 10 mins (see image by Dr. David Bertwistle).
WB (7)
1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa). Multiple bands with this antibody have frequently been reported. We believe that this is due to the different modification states of p19ARF. Nuclear extraction may be necessary in some samples.
IHC-P (2)
Use at an assay dependent concentration. PubMed: 11726500
说明
ICC/IF
Use at an assay dependent concentration.
IHC-Fr
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC
Use a concentration of 1 µg/ml. Fix with a 1:1 mixture of methanol:acetone at -20C for 10 mins (see image by Dr. David Bertwistle).
WB
1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa). Multiple bands with this antibody have frequently been reported. We believe that this is due to the different modification states of p19ARF. Nuclear extraction may be necessary in some samples.
IHC-P
Use at an assay dependent concentration. PubMed: 11726500

靶标

  • 相关性

    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.
  • 细胞定位

    Nuclear
  • 数据库链接

    • Entrez Gene: 12578 Mouse
    • SwissProt: Q64364 Mouse
    • 别名

      • ARF antibody
      • CDKN2A antibody
      • Cyclin dependent kinase inhibitor 2A antibody
      • INK4 antibody
      • MTS1 antibody
      • P19 antibody
      see all

    图片

    • Western blot - Anti-CDKN2A/p19ARF antibody (ab80)
      Western blot - Anti-CDKN2A/p19ARF antibody (ab80)This image is courtesy of Charles Sherr
      All lanes : Anti-CDKN2A/p19ARF antibody (ab80) at 1 µg/ml

      Lanes 1-2 : mouse embryonic fibroblast total cell extract.
      Lane 3 : mouse embryonic fibroblast total cell extract, with the p19ARF gene knocked out as a negative control.

      Lysates/proteins at 30 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 17 kDa



      Western blot of primary Mouse Embryo Fibroplasts (MEFs) using ab80 at 1 µg/ml.  (30 µg of protein from total cell extracts per lane.)

    • Immunocytochemistry - Anti-CDKN2A/p19ARF antibody (ab80)
      Immunocytochemistry - Anti-CDKN2A/p19ARF antibody (ab80)Pictures were generously provided by Dr. David Bertwistle

      ab80 antibody was tested on NIH 3T3 cells (which have a deletion of the p19ARF gene) as a negative control (-) and NIH 3T3 cells engineered to express HA-tagged p19ARF as a positive control (+).

      Cells grown on coverslips were fixed with a 1:1 mixture of methanol:acetone at -20C for 10 mins. After drying, rehydrating and blocking, the co

    实验方案

    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    数据表及文件

    • SDS download

    • Datasheet download

      Download

    文献 (212)

    发表研究结果有使用 ab80?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab80 被引用在 212 文献中.

    • Bazzar W  et al. Pharmacological inactivation of CDK2 inhibits MYC/BCL-XL-driven leukemia in vivo through induction of cellular senescence. Cell Cycle 20:23-38 (2021). PubMed: 33356836
    • Chen P  et al. Intramuscular injection of Botox causes tendon atrophy by induction of senescence of tendon-derived stem cells. Stem Cell Res Ther 12:38 (2021). PubMed: 33413592
    • Li Y  et al. Stabilization of p18 by deubiquitylase CYLD is pivotal for cell cycle progression and viral replication. NPJ Precis Oncol 5:14 (2021). PubMed: 33654169
    • Kotowski K  et al. 3PO as a Selective Inhibitor of 6-Phosphofructo-2-Kinase/Fructose-2,6-Biphosphatase 3 in A375 Human Melanoma Cells. Anticancer Res 40:2613-2625 (2020). PubMed: 32366406
    • Mikawa R  et al. p19Arf Exacerbates Cigarette Smoke-Induced Pulmonary Dysfunction. Biomolecules 10:N/A (2020). PubMed: 32192082
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
    提交评价 提交问题

    1-10 of 39 Abreviews or Q&A

    Western blot abreview for Anti-CDKN2A/p19ARF antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Liver)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (10)
    Loading amount
    20 µg
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5%
    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.

    Abcam user community

    Verified customer

    提交于 May 30 2022

    Western blot abreview for Anti-CDKN2A/p19ARF antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Liver)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (15%)
    Loading amount
    30 µg
    Treatment
    200µM Palmitate
    Specification
    Liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    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.

    Abcam user community

    Verified customer

    提交于 Jul 08 2020

    Western blot abreview for Anti-CDKN2A/p19ARF antibody

    Good
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (lung cancer)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    lung cancer
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    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.

    Abcam user community

    Verified customer

    提交于 Aug 20 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-CDKN2A/p19ARF antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (HF)
    Permeabilization
    Yes - PBS-TWEEN 0.05%
    Specification
    HF
    Blocking step
    Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Acetone
    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.

    Abcam user community

    Verified customer

    提交于 Feb 23 2018

    IHC - Wholemount

    Average
    Abreviews
    Abreviews
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (inner ear tissue)
    Specification
    inner ear tissue
    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.

    Abcam user community

    Verified customer

    提交于 Jun 11 2013

    Western blot abreview for Anti-CDKN2A/p19ARF antibody

    Excellent
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse whole cell lysate)
    Loading amount
    30 µg
    Specification
    mouse whole cell lysate
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    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.

    Abcam user community

    Verified customer

    提交于 Apr 05 2013

    Question


    Inquiry: Hi, I am having trouble visualizing p19ARF on a WB. I have a band at ˜30 KDa, but none near 19 KDa. I loaded 20ug mouse thymus lysate. A 12% gel was run and then I did a wet transfer onto a nitrocellulose membrane for 2 hours. The WB was blocked for 1h in 5% milk at RT. I incubated with the primary antibody at 4C O/N in 5% milk (1:1000 dilution) and revealed with blot with anti-Rabbit-HRP antibodies which work for other samples. All washes were done with 0.1% TBS-T. Thank you.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 08 2012

    Answer

    Thank you for contacting us. Depending on the gel, samples, and buffer, the bands can appear 10 kD from the "predicted" size on your gel. Since the concentration of your antibody is 0.6 mg/ml and we suggest using 1 ug/ml on your blot, you could try using 1/500 primary overnight to see if any other bands appear. You could also use the blocking peptide ab2659 to see if the band on your blot is specifically blocked.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. We do guarantee our products to work as specified on the datasheet and can replace, credit or refund the product within 6 months if you are not satisfied with your results.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Nov 08 2012

    Question

    On the basis of the result of microarray , it is reasonable that the upper band means p19 expression.

    But the positive control (NIH3T3) located on the lower band. Moreover , the band size seems to be a little bit lower than 19 kDa.

    I don’t know which band shows the actual p19. Also, is there any possibility that the location of the band is different between samples and NIH3T3 ?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 30 2012

    Answer

    Thank you for contacting us.

    Based on this image alone it seems like the lower band is your CDKN2A/p19ARF band. It is possible for band size to change slightly due to sample preparation or buffer, however usually the positive control will be a good indicator of where the protein of interest is. One strategy to confirm the band might be to use the blocking peptide (the peptide used as the immunogen) to specifically block the CDKN2A/p19ARF protein. We have this available as ab
    2659.


    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    回复于 Oct 30 2012

    Question

    Hello,

    Yesterday, I used your p19ARF antibody(ab80) for my samples. Also I set NIH3T3 whole cell lysate as a positive control, but it didn’t work. (no band.)

    I borrowed this cell line from other lab. After I cultured, I extracted lysate by myself.

    I think there is some possibility that my NIH3T3 had any problems. So, I plan to order your cell lysate and check some mouse embryonic fibroblast cell line; ab7179,ab14874,ab7901,ab3874.

    I’d like to ask you which is the best as positive control for p19ARF antibody? Or is there no difference among them?

    Thank you,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 11 2012

    Answer

    Thank you very much for contacting us with your enquiry.

    I'm sorry to hear that there were no bands in Western blot using this antibody. I would expect any of the MEF cell lines to work as a positive control, but I suggest using a whole cell lysate (ab14874 is a nuclear fraction) since that is what we have tested with ab80. What kind of samples are you using other than the MEF lysate?

    If you'd like to send some more details about your protocol ( how much protein was loaded, dilution of antibody, blocking solution, etc) I'd be happy to take a look and see if we have any suggestions to improve the results.

    I look forward to hearing from you. If there's anything else that we can do for you, please let me know and I'll be happy to help.

    Read More

    Abcam Scientific Support

    回复于 Oct 11 2012

    Question

    This antibody is not showing any bands in WB.  I would like a replacement with a new lot.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 05 2011

    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab80 from a different lot. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.  

    Read More

    Abcam Scientific Support

    回复于 Dec 05 2011

    1-10 of 39 Abreviews or Q&A

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