Product nameAnti-CDKN2A/p19ARF antibody
See all CDKN2A/p19ARF primary antibodies
DescriptionRabbit polyclonal to CDKN2A/p19ARF
SpecificityOn 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 applicationsSuitable for: ICC/IF, ICC, ELISA, IHC-P, WB, IHC-Frmore details
Species reactivityReacts with: Mouse
Does not react with: Rat, Human
- mouse embryonic fibroblasts lysate
General notesThe blocking peptide for this antibody is available (ab2659)
Storage instructionsShipped 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.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
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...
PurityImmunogen affinity purified
Purification notesThe 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.
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||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). 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.|
RelevanceThe 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.
- ARF antibody
- CDKN2A antibody
- Cyclin dependent kinase inhibitor 2A antibody
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.
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.
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 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) »