The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesIF: 1/1000.
IHC-F: Use at a concentration of 5 µg/ml.
IP: Use at an assay dependent dilution.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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.
Immunoprecipitation using anti-p19ARF 5C3. NIH3T3 cells expressing HA-p19ARF were incubated with a control rat IgG (lane 1) or anti-p19ARF 5C3 (2). Precipitated proteins were analysed by Western blot using anti-HA antibody.
Immunofluorescent detection of p19ARF. MEF cells (1 and 2) or NIH3T3 cells with arf gene deleted (3 and 4) were stained with anti-p19ARF 5C3 followed by a fluorescent-labelled anti-rat IgG (1 and 3) and Hoechst dye to visualise nuclei (2 and 4).
References for Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)
This product has been referenced in:
Bartesaghi S et al. Inhibition of oxidative metabolism leads to p53 genetic inactivation and transformation in neural stem cells. Proc Natl Acad Sci U S A112:1059-64 (2015).
Read more (PubMed: 25583481) »
Miceli AP et al. Hypergrowth mTORC1 signals translationally activate the ARF tumor suppressor checkpoint. Mol Cell Biol32:348-64 (2012).
Read more (PubMed: 22064482) »