Overview

  • Product name
    Anti-CDKN2A/p19ARF antibody
    See all CDKN2A/p19ARF primary antibodies
  • Description
    Rabbit polyclonal to CDKN2A/p19ARF
  • Host species
    Rabbit
  • Specificity
    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.
  • Tested applications
    Suitable for: ICC/IF, ICC, ELISA, IHC-P, WB, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse
    Does not react with: Rat, Human
  • Immunogen

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

    (Peptide available as ab2659.)

  • Positive control
    • mouse embryonic fibroblasts lysate
  • General notes
    The blocking peptide for this antibody is available (ab2659)

Properties

Applications

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.

Application Abreviews Notes
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.

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
    Nuclear
  • Database links
  • Alternative names
    • ARF antibody
    • CDKN2A antibody
    • Cyclin dependent kinase inhibitor 2A antibody
    • INK4 antibody
    • MTS1 antibody
    • P19 antibody
    see all

Images

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

    Lanes 1-2 : 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?)



    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

References

This product has been referenced in:
  • Oldrini B  et al. Somatic genome editing with the RCAS-TVA-CRISPR-Cas9 system for precision tumor modeling. Nat Commun 9:1466 (2018). Read more (PubMed: 29654229) »
  • Francis JC  et al. SOX9 is a driver of aggressive prostate cancer by promoting invasion, cell fate and cytoskeleton alterations and epithelial to mesenchymal transition. Oncotarget 9:7604-7615 (2018). Read more (PubMed: 29484136) »

See all 188 Publications for this product

Customer reviews and Q&As

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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
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Abcam user community

Verified customer

Submitted Feb 23 2018

Abreviews
Application
IHC - Wholemount
Sample
Mouse Tissue (inner ear tissue)
Specification
inner ear tissue
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Verified customer

Submitted 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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Submitted Apr 05 2013

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
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Submitted 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
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Submitted 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
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Submitted Jul 09 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (pancreatic tumor)
Loading amount
50 µg
Specification
pancreatic tumor
Gel Running Conditions
Non-reduced Denaturing (15%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
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Submitted Jun 10 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
3% BSA + 2% NGS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
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Mrs. Demet Caglayan

Verified customer

Submitted Nov 15 2007

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 (Kidney (glomerulus))
Specification
Kidney (glomerulus)
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization
No
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: 22°C
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Verified customer

Submitted Sep 18 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Murine embryonic fibroblast)
Loading amount
30 µg
Specification
Murine embryonic fibroblast
Treatment
oncogene-transduced primary murine cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Dr. Jose A Costoya

Verified customer

Submitted Mar 07 2007

1-10 of 12 Abreviews

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