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Product name
Anti-CDKN2A/p19ARF antibody
See all CDKN2A/p19ARF products (4) ...
Description
Rabbit polyclonal to CDKN2A/p19ARF
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
ICC/IF, ICC, ELISA, IHC-P, WB, IHC-Frmore details
Cross 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
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: 1% BSA
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
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.
Clonality
Polyclonal
Isotype
IgG
Research areas
Cancer >> Oncoproteins/suppressors >> Tumor suppressors >> INK4a/Arf
Cancer >> Oncoproteins/suppressors >> Tumor suppressors >> p53 pathway
Cancer >> Cell cycle >> Cell cycle inhibitors >> Ink4
Epigenetics and Nuclear Signaling >> Cell cycle >> Cell Cycle Inhibitors >> Ink4
Epigenetics and Nuclear Signaling >> Transcription >> Cancer susceptibility >> Tumor Suppressors
Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Ink4
Applications
Show applications key
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.
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 dilution.
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 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
Alternative names
- MTS1 antibody
- ARF antibody
- CDK4 inhibitor P16 INK4 antibody
see all
Database links
- Entrez Gene: 12578 Mouse
- SwissProt: Q64364 Mouse
- Unigene: 4733 Mouse
Anti-CDKN2A/p19ARF antibody images:
Western blot - p19ARF antibody (ab80)
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?)
Charles Sherr
Immunocytochemistry - p19ARF antibody (ab80)
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 coverslips were incubated with ab80 at a concentration of 1ug/ml, diluted in PBS for 1 hour at room temperature. The antibody was then visualised using standard anti-rabbit secondary antibodies coupled to Texas Red.
Pictures were generously provided by Dr. David Bertwistle
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - p19ARF antibody (ab80)
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.
This image is courtesy of an anonymous Abreview
Immunocytochemistry/ Immunofluorescence - p19ARF antibody (ab80)
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.
This image is courtesy of an Abreview submitted by Mrs Nanna Forsberg
Immunohistochemistry (Frozen sections) - CDKN2A/p19ARF antibody (ab80)
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.
This image is courtesy of an anonymous Abreview
References for Anti-CDKN2A/p19ARF antibody (ab80)
This product has been referenced in:
- Carneiro C et al. p27 deficiency desensitizes Rb-/- cells to signals that trigger apoptosis during pituitary tumor development. Oncogene 22:361-9 (2003).Read more (PubMed: 12545157) »
- Peeper DS et al. A functional screen identifies hDRIL1 as an oncogene that rescues RAS-induced senescence. Nat Cell Biol 4:148-53 (2002).Read more (PubMed: 11812999) »
See all 106 publications for this product
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"