Immunofluorescent analysis of p14ARF/p16beta (green) showing staining in the nucleus of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a p14ARF/p16beta monoclonal antibody (Product # MA5-14260) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human|
|Published species reactivity||Fruit fly, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant p14ARF protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||2 µg/ml|
|Western Blot (WB)||2-4 µg/ml|
* 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.
MA5-14260 targets p14ARF/p16beta in ICC/IF, IP, and WB applications and shows reactivity with Human samples.
The MA5-14260 immunogen is recombinant p14ARF protein.
The INK4a-ARF locus encodes two unrelated proteins both of which function in tumor suppression. p14ARF arrests the cell cycle in a p53-dependent manner. p14ARF binds to mdm2 and promotes the rapid degradation of mdm2 proteinrequired for mdm2 modification and concurrent p53 stabilization and accumulation. This interaction is mediated by the exon 1beta-encoded N-terminal domain of p14ARF and a C-terminal region of mdm2. Deletion of the ARF-INK4a locus simultaneously impairs both the INK4a-cyclin D/cdk4-RB and the ARF-mdm2-p53 pathways.
IP-MS enrichment of CDKN2A (LFQ intensity): CDKN2A was enriched 98-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. MA5-14260). 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A patient with a noninvasive mucinous ovarian borderline tumor presenting with late pleural metastases.
MA5-14260 was used in immunohistochemistry - paraffin section to describe a patient diagnosed with a noninvasive intestinal-type mucinous ovarian borderline tumor presenting with pleural metastases
|Simons M,Nagtegaal ID,Overbeek LI,Flucke U,Massuger LF,Bulten J||International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists (34:143)||2015|
ARF represses androgen receptor transactivation in prostate cancer.
MA5-14260 was used in immunohistochemistry, immunoprecipitation, and western blot to study the role of p14ARP as an androgen receptor-interacting protein that blocks its transactivation in prostate cancer cells
|Lu W,Xie Y,Ma Y,Matusik RJ,Chen Z||Molecular endocrinology (Baltimore, Md.) (27:635)||2013|
|Not Applicable||Not Cited||
Mimicking p14ARF phosphorylation influences its ability to restrain cell proliferation.
MA5-14260 was used in immunoprecipitation and western blot to report that p14ARF is a PKC target
|Vivo M,Ranieri M,Sansone F,Santoriello C,Calogero RA,Calabrò V,Pollice A,La Mantia G||PloS one (8:null)||2013|
Myc-ARF (alternate reading frame) interaction inhibits the functions of Myc.
MA5-14260 was used in immunoprecipitation to investigate the interaction of ARF with c-Myc and its influence on c-Myc function
|Datta A,Nag A,Pan W,Hay N,Gartel AL,Colamonici O,Mori Y,Raychaudhuri P||The Journal of biological chemistry (279:36698)||2004|
p16INK4A-silencing augments DNA damage-induced apoptosis in cervical cancer cells.
MA5-14260 was used in western blot to study the role of p16INK4A-silencing in DNA damage-induced apoptosis in cervical cancer cells
|Lau WM,Ho TH,Hui KM||Oncogene (26:6050)||2007|
|Fruit fly||Not Cited||
Functional identification of Api5 as a suppressor of E2F-dependent apoptosis in vivo.
MA5-14260 was used in western blot to investigate the effect of Api5 on the E2F-dependent apoptosis in vivo
|Morris EJ,Michaud WA,Ji JY,Moon NS,Rocco JW,Dyson NJ||PLoS genetics (2:null)||2006|
Bak functionally complements for loss of Bax during p14ARF-induced mitochondrial apoptosis in human cancer cells.
MA5-14260 was used in western blot to study the ability of Bak to functionally complement for loss of Bax during mitochondrial apoptosis in human cancer cells
|Hemmati PG,Güner D,Gillissen B,Wendt J,von Haefen C,Chinnadurai G,Dörken B,Daniel PT||Oncogene (25:6582)||2006|
Antiviral action of the tumor suppressor ARF.
MA5-14260 was used in western blot to study the role of the tumor suppressor ARF in viral infection surveillance
|García MA,Collado M,Muñoz-Fontela C,Matheu A,Marcos-Villar L,Arroyo J,Esteban M,Serrano M,Rivas C||The EMBO journal (25:4284)||2006|
p14ARF induces G2 cell cycle arrest in p53- and p21-deficient cells by down-regulating p34cdc2 kinase activity.
MA5-14260 was used in western blot to study the mechanism by which p14ARF induces G2 cell cycle arrest in p53- and p21-deficient cells
|Normand G,Hemmati PG,Verdoodt B,von Haefen C,Wendt J,Güner D,May E,Dörken B,Daniel PT||The Journal of biological chemistry (280:7118)||2005|
Different requirements for the cytostatic and apoptotic effects of type I interferons. Induction of apoptosis requires ARF but not p53 in osteosarcoma cell lines.
MA5-14260 was used in western blot to investigate the mechanism for the effect of type I interferons on cell death
|Sandoval R,Xue J,Pilkinton M,Salvi D,Kiyokawa H,Colamonici OR||The Journal of biological chemistry (279:32275)||2004|
Immortalization of human mammary epithelial cells is associated with inactivation of the p14ARF-p53 pathway.
MA5-14260 was used in western blot to study the role of p14ARF-p53 pathway inactivation in the immortalization of human mammary epithelial cells
|Shamanin VA,Androphy EJ||Molecular and cellular biology (24:2144)||2004|
p14ARF silencing by promoter hypermethylation mediates abnormal intracellular localization of MDM2.
MA5-14260 was used in western blot to study the effects of epigenetic silencing of p14ARF on the subcellular localization of MDM2
|Esteller M,Cordon-Cardo C,Corn PG,Meltzer SJ,Pohar KS,Watkins DN,Capella G,Peinado MA,Matias-Guiu X,Prat J,Baylin SB,Herman JG||Cancer research (61:2816)||2001|
p14ARF, a prognostic predictor in HPV-negative vulvar carcinoma.
MA5-14260 was used in immunohistochemistry to study the prognostic value of p14ARF in HPV-negative vulvar carcinoma
|Knopp S,Nesland JM,Tropé C,Holm R||American journal of clinical pathology (126:266)||2006|
Vulvar squamous cell carcinoma is a multifactorial disease following two separate and independent pathways.
MA5-14260 was used in immunohistochemistry to study the pathways leading to vulvar squamous cell carcinoma
|van der Avoort IA,Shirango H,Hoevenaars BM,Grefte JM,de Hullu JA,de Wilde PC,Bulten J,Melchers WJ,Massuger LF||International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists (25:22)||2006|
p14ARF expression in invasive breast cancers and ductal carcinoma in situ--relationships to p53 and Hdm2.
MA5-14260 was used in immunohistochemistry to study the expression and cellular distribution of p14ARF, p53 and Hdm2 in invasive breast cancers with ductal carcinoma in situ
|Vestey SB,Sen C,Calder CJ,Perks CM,Pignatelli M,Winters ZE||Breast cancer research : BCR (6:R571)||2004|
Aberrant expression of tumor suppressor proteins in the Ewing family of tumors.
MA5-14260 was used in immunohistochemistry to investigate tumor suppressor proteins in the Ewing family of tumors
|Maitra A,Roberts H,Weinberg AG,Geradts J||Archives of pathology and laboratory medicine (125:1207)||2001|
Ebp1 is a dsRNA-binding protein associated with ribosomes that modulates eIF2alpha phosphorylation.
MA5-14260 was used in immunocytochemistry to characterize the localization of Ebp1 and its function
|Squatrito M,Mancino M,Sala L,Draetta GF||Biochemical and biophysical research communications (344:859)||2006|
DNA damage disrupts the p14ARF-B23(nucleophosmin) interaction and triggers a transient subnuclear redistribution of p14ARF.
MA5-14260 was used in immunocytochemistry to investigate the impact of DNA damage on subnuclear distribution of ARF and its interaction with B23 in human tumor cells
|Lee C,Smith BA,Bandyopadhyay K,Gjerset RA||Cancer research (65:9834)||2005|
EBP1 is a nucleolar growth-regulating protein that is part of pre-ribosomal ribonucleoprotein complexes.
MA5-14260 was used in immunocytochemistry to investigate the cellular function of EBP1
|Squatrito M,Mancino M,Donzelli M,Areces LB,Draetta GF||Oncogene (23:4454)||2004|
Nitric oxide induces phosphorylation of p53 and impairs nuclear export.
MA5-14260 was used in immunocytochemistry to study the role of nitric oxide in inducing the phosphorylation and impaired nuclear export of p53
|Schneiderhan N,Budde A,Zhang Y,Brüne B||Oncogene (22:2857)||2003|
A melanoma-predisposing germline CDKN2A mutation with functional significance for both p16 and p14ARF.
MA5-14260 was used in immunocytochemistry to investigate the effect of melanoma-predisposing germline CDKN2A mutation on p16 and p14ARF activation
|Hashemi J,Lindström MS,Asker C,Platz A,Hansson J,Wiman KG||Cancer letters (180:211)||2002|