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
  • 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, ICC, ELISA, IHC-P, WB, IHC-Frmore details
  • 种属反应性
    与反应: Mouse
    不与反应: Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Mouse p19ARF.

    (Peptide available as ab2659.)

  • 阳性对照
    • mouse embryonic fibroblasts lysate
  • 常规说明The blocking peptide for this antibody is available (ab2659)

性能

应用

Our Abpromise guarantee covers the use of ab80 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/250.
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).
ELISA Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. PubMed: 11726500
WB 1/1000. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa).Can be blocked with Mouse CDKN2A/p19ARF peptide (ab2659). 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-Fr 1/100.

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

Anti-CDKN2A/p19ARF antibody 图像

  • All lanes : Anti-CDKN2A/p19ARF antibody (ab80) at 1 µg/ml

    Lane 1 : 30 ug of mouse embryonic fibroblast total cell extract.
    Lane 2 : 30 ug of mouse embryonic fibroblast total cell extract.
    Lane 3 : 30 ug of mouse embryonic fibroblast total cell extract, with the p19ARF gene knocked out as a negative control.


    Performed under reducing conditions.

    Predicted band size : 17 kDa
    Observed band size : 19 kDa (why is the actual band size different from the predicted?)

    This image is courtesy of Charles Sherr

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

  • 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

  • ab80 at 1/300 staining mouse kidney (glomerulus) tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 45 minutes. An HRP conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • ab80 at 1/300 staining mouse brain cells by ICC/IF.

    Paraformaldehyde fixation, Triton X-100 permeabilization, followed by blocking with 3% BSA + 2% NGS for 1 hour at 20°C.

    Incubation with the primary antibody was carried out for 1 hour at 20°C.  The secondary antibody was an Alexa Fluor® conjugated Donkey anti-Rabbit polyclonal, diluted 1/400.

    See Abreview

  • ab80 staining CDKN2A/p19ARF in Mouse pancreatic neoplastic tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS) for 8 hours at 4°C. An Alexa Fluor®555-conjugated Donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody. Nuclei were stained by DAPI.

    See Abreview

Anti-CDKN2A/p19ARF antibody (ab80)参考文献

This product has been referenced in:
  • Lu Y  et al. CDK4 deficiency promotes genomic instability and enhances Myc-driven lymphomagenesis. J Clin Invest 124:1672-84 (2014). WB . Read more (PubMed: 24614102) »
  • Kanatsu-Shinohara M  et al. Skp1-Cullin-F-box (SCF)-type ubiquitin ligase FBXW7 negatively regulates spermatogonial stem cell self-renewal. Proc Natl Acad Sci U S A 111:8826-31 (2014). IHC-Fr ; Mouse . Read more (PubMed: 24879440) »

See all 122 Publications for this product

Product Wall

Abreviews
Application IHC - Wholemount
Sample Mouse Tissue (inner ear tissue)
Specification inner ear tissue
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Abcam user community

Verified customer

提交于 Jun 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Abcam user community

Verified customer

提交于 Apr 05 2013

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 cou...

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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...

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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 ...

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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 ca...

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My colleague from US Customer Service has passed your message to me. Thank you for your recent call regarding ab80. I am very sorry to hear that you are having problems with this antibody. All our customer feedback, including complaints are monit...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (mouse pancreatic cancer primary)
Specification mouse pancreatic cancer primary
Fixative Acetone
Permeabilization Yes - PBS + Triton 0.025%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jul 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Mouse skin papilloma)
Specification Mouse skin papilloma
Fixative Paraformaldehyde
Antigen retrieval step None - Buffer/Enzyme Used: citrate
Permeabilization Yes - PBS+Triton X-100 0.025%
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (mouse pancreatic neoplasia)
Specification mouse pancreatic neoplasia
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Jul 09 2010

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