p14ARF/CDKN2A Antibody

Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - ARF expression levels in OCI/AML3 cells after proteasome treatment. This image was submitted via customer Review.

Immunocytochemistry/Immunofluorescence: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested in HeLa cells with Dylight 488 (green). Nuclei were counterstained with DAPI (blue). Tubulin was stained with alpha tubulin (red).

Flow Cytometry: p14ARF/CDKN2A Antibody [NB200-111] - p14ARF antibody was tested at 1:400 in HeLa cells using an Alexa Fluor 488 secondary (shown in purple). M1 is defined by unstained cells.

Western Blot: p14ARF/CDKN2A Antibody [NB200-111] - WB analysis of HeLa Whole Cell Lysate (NB800-PC1) using p14ARF antibody (lot C) employing ECL detection method.

• Reactivity: Hu, Mu
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1.0 mg/ml

p14ARF/CDKN2A Antibody Summary

• Immunogen: A synthetic peptide made to a portion of human p14ARF (between residues 50-150). [Swiss-Prot# Q8N726]
• Localization: Nuclear
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: CDKN2A
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot
  • ELISA 1:100-1:2000
  • Flow Cytometry 1:400
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:10-1:500
  • Immunohistochemistry-Frozen 1:10-1:500
  • Immunohistochemistry-Paraffin 1:10-1:500
  • Immunoprecipitation
• Application Notes: This p14ARF antibody is used for Western blotting, where a band is seen at where a band is seen at ~16 kDa, representing p14ARF (p14ARF tends to run slightly higher than the theoretical MW of 14 kDa). Additional faint bands may be seen at ~32 and 47 kDa. WB application of this product is mentioned in publication with PMID: 21636682. In ICC/IF, nuclear focal staining was observed in HeLa cells.
• Positive Control:

  p16 Lysate (NBL1-09052)
  p14ARF/CDKN2A Lysate (NBP2-66410)

Reactivity Notes

NB 200-111 reacts with human p14 ARF.

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.05% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for p14ARF/CDKN2A Antibody

• ARF
• CDK4 inhibitor p16-INK4
• CDK4I
• CDK4IP14ARF
• CDKN2
• CDKN2A
• cell cycle negative regulator beta
• CMM2P16-INK4A
• Cyclin-dependent kinase 4 inhibitor A
• cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
• cyclin-dependent kinase inhibitor 2A
• INK4
• INK4a
• MLM
• MLMP16INK4
• MTS-1
• MTS1P14
• Multiple tumor suppressor 1
• p14
• p14ARF
• p16
• p16-INK4
• p16INK4a
• p16-INK4a
• P19
• p19ARF
• TP16

Background

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 1 alpha) 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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for p14ARF/CDKN2A Antibody (NB200-111)(16)

Publications using NB200-111

• Crawford Parks TE, Marcellus KA, Langill J et al. Novel Roles for Staufen1 in Embryonal and Alveolar Rhabdomyosarcoma via c-myc-dependent and -independent events. Sci Rep. 2017 Feb 17 [PMID: 28211476] (WB, Human)
• Ghosh M, Ryan RO. Curcumin homing to the nucleolus: mechanism for initiation of an apoptotic program. J Nutr. Biochem. 2014 Aug 01 [PMID: 25172633] (WB, Human)
• Hamilton Garth, Abraham Aswin G, Morton Jennifer et al. AKT regulates NPM dependent ARF localization and p53mut stability in tumors. Oncotarget. 2014 Aug 15 [PMID: 25071014] (ICC/IF)
• Williams RT, Barnhill LM, Kuo HH et al. Chimeras of p14ARF and p16: Functional Hybrids with the Ability to Arrest Growth. PLoS ONE 2014 Feb 7 [PMID: 24505435] (WB, Human)
• Chio II, Sasaki M, Ghazarian D, Moreno J, Done S, Ueda T, Inoue S, Chang YL, Chen NJ, Mak TW. TRADD contributes to tumour suppression by regulating ULF-dependent p19(Arf) ubiquitylation. Nat Cell Biol;14(6):625-33. 2012 May 6. [PMID: 22561347] (WB, Mouse, Human)
• Muniz V, Barnes JM, Paliwal S et al. The ARF tumor suppressor inhibits tumor cell colonization independent of p53 in a novel mouse model of pancreatic ductal adenocarcinoma metastasis. Mol Cancer Res 9(7):867-77. 2011 Jul. [PMID: 21636682] (WB, Mouse)
• Chen D, Yoon JB, Gu W. Reactivating the ARF-p53 axis in AML cells by targeting ULF. Cell Cycle 2010 Aug 1 [PMID: 20699639] (IP, Human)
• Greiner T. Who chooses the leaders of UN organisations? Lancet 2010 Mar 27 [PMID: 20346814] (Human)
• Chen D, Shan J, Zhu WG, Qin J, Gu Wet al. Transcription-independent ARF regulation in oncogenic stress-mediated p53 responses. Nature 2010 Mar 25 [PMID: 20208519] (IP, Human)
• Wang W-H, Hullinger RL, risani OM. Hepatitis B virus X protein via the p38MAPK pathway induces E2F1 release and ATR kinase activation mediating p53 apoptosis. J Biol Chem 283(37):25455-67. 2008 Sep 12. [PMID: 18606816] (WB, Mouse)
• Freedberg DE et al. Frequent p16-independent inactivation of p14ARF in human melanoma. J Natl Cancer Inst 100(11):784-95. 2008 Jun 4. [PMID: 18505964] (IHC, Human)
• Mascaux, C et al. The role of NPM, p14arf and MDM2 in precursors of bronchial squamous cell carcinoma. Eur Respir J 32(3):678-86. 2008 Sep. [PMID: 18480108] (IHC, Human)
• Miller SJ et al. Large-scale molecular comparison of human schwann cells to malignant peripheral nerve sheath tumor cell lines and tissues. Cancer Res 66(5):2584-91. 2006 Mar 1. [PMID: 16510576] (WB, Human)
• Kuo, ML et al. N-terminal polyubiquitination and degradation of the Arf tumor suppressor. Genes Dev 18(15):1862-74. 2004 Aug 1. [PMID: 15289458] (WB, Human, Mouse)
• Magro PG, Russo AJ, Li WW et al. p14ARF expression increases dihydrofolate reductase degradation and paradoxically results in resistance to folate antagonists in cells with nonfunctional p53. Cancer Res 2004 Jun 15 [PMID: 15205349] (Human)
• Fatyol, K et al. The p14ARF tumor suppressor protein facilitates nucleolar sequestration of hypoxia-inducible factor-1alpha (HIF-1alpha ) and inhibits HIF-1-mediated transcription. J Biol Chem 276(30):28421-9. 2001 Jul 27. [PMID: 11382768] (WB, Human, Mouse)

Reviews for p14ARF/CDKN2A Antibody (NB200-111) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 1 species: Human.

reviewed by: Ming Hua WB Human 09/01/2017

Summary

• Application: Western Blot
• Sample Tested: OCI/AML3 cells,human AML NPM cytoplasmic mutant cell line
• Species: Human
• Lot: H-1

reviewed by: Ronald Miller WB Human 05/29/2014

Summary

• Application: Western Blot
• Sample Tested: Young and senescent cells
• Species: Human
• Comments: Its difficult to detect this protein because of low abundance in cells. A longer exposure helps capture the signal.

Blocking

• Blocking Details: 5% BSA in 1X PBS

Primary Anitbody

• Dilution Ratio: 1:1000 in 1X PBS with troton X-100 (0.1%) at 4 degree O/N

Secondary Antibody

• Secondary Description: Goat anti rabbit-HRP conjugated
• Secondary Manufacturer Cat#: 65-6120
• Secondary Concentration: 1:20,000

Details

• Detection Notes: ECL, 5 min exposure with film

Comments

• Comments: Its difficult to detect this protein because of low abundance in cells. A longer exposure helps capture the signal.

FAQs for p14ARF/CDKN2A Antibody (NB200-111). (Showing 1 - 2 of 2 FAQs).

1. While looking for antibodies targeting human p14ARF, I found a question: Based on the information on UniProt, human p14ARF has the length of only 132 a.a. However, the immunogen of NB200-159 is described as... between residue 125 and the C-terminus (residue 173)...,...between residues 100-173... for NB110-59085, and...between residues 50-150... for NB200-111.
   • Since these antibodies were produced, UniProt has since truncated the protein sequence. These antibodies are still directed towards p14ARF and the immunogens will be updated on our website to the updated amino acid ranges. Since you won't be able to see this until this weekend or early next week, the immunogen for NB200-111 falls between amino acids 50 and 132. The immunogen for NB200-159 falls between amino acids 90 and 132 and the immunogen for NB110-59085 falls between amino acids 50 and 132.
2. Do you measure the isotype purity of NB200-111? I realize this is antigen affinity purified, and will be almost completely IgG, but do you measure for IgA/M amount by chance?
   • We unfortunately do not measure isotype purity for our products.

Bioinformatics

• Gene Symbol: CDKN2A
• Entrez:
  • 1029(Human)
  • 12578(Mouse)
  • 25163(Rat)
• Uniprot:
  • Q8N726(Human)