Immunocytochemical analysis of p14ARF Antibody in HeLa cells. Panel one is a brightfield image, panel two shows the immunofluorescence staining of p14RF detected using Dylight 488 and panel three is an overlay of showing the nuclear localization of the p14ARF staining.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide made to a portion of human p14ARF (between residues 50-150) .|
|Purification||Antigen affinity chromatography|
|Contains||0.05% sodium azide|
|Storage Conditions||-20°C or -80°C if preferred|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:400|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:500|
|Western Blot (WB)||1:100-1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
p14ARF tends to run slightly higher than the theoretical MW of 14 kDa.
Suggested positive control: BT549 cells or HeLa whole cell extract, antigen standard for CDKN2A (transient overexpression lysate).
The INK4a-ARF locus is comprised of two tumor suppressors, p16INK4a and p14ARF. These two proteins are encoded through differential splicing of alternative first exons. The p16INK4a (exon 1alpha) protein inhibits the cyclin D-dependent kinases (CDK) that control the phosphorylation of the Rb protein and cell proliferation. The p14ARF gene product complexes with the MDM2 protein within the nucleus, thus modulating the activity of the p53 protein. P14ARF is a potent tumor suppressor in the presence of wild-type p53, while mutant p53 substantially reduces growth inhibition by p14ARF.
IP-MS enrichment of CDKN2A (LFQ intensity): CDKN2A was enriched 1449-fold from BT549 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and CDKN2A antibody (Part No. PA1-16639). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
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