Recombinant Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
![Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-311236-anti-cdkn2a-p14arf-antibody-epr17878-western-blot.jpg "Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF
- Suitable for: Flow Cyt (Intra), ICC/IF, IP, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CDKN2A/p14ARF antibody [EPR17878]
See all CDKN2A/p14ARF primary antibodies -
Description
Rabbit monoclonal [EPR17878] to CDKN2A/p14ARF -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa whole cell lysate transfected with CDKN2A/p14ARF with His-tag; PC-3 and HEK-293 whole cell lysates. ICC/IF: HeLa cells transfected with CDKN2A/p14ARF. Flow Cyt (intra): HeLa cells transfected with CDKN2A/p14ARF-GFP. IP: CDKN2A/p14ARF transfected HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17878 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185620 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt (Intra) |
1/120.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| ICC/IF |
1/250.
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| IP |
1/80.
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| WB |
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa).
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| Notes |
|---|
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Flow Cyt (Intra)
1/120. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF
1/250. |
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IP
1/80. |
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WB
1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 14 kDa). |
Target
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Relevance
The 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. -
Cellular localization
Cytoplasmic and Nuclear -
Database links
- Entrez Gene: 1029 Human
- Omim: 600160 Human
- SwissProt: P42771 Human
- SwissProt: Q8N726 Human
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Alternative names
- ARF antibody
- CDK4 inhibitor p16 INK4 antibody
- CDK4I antibody
see all
Images
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Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/1000 dilution
Lane 1 : Wild-type HeLa whole cell lysate
Lane 2 : CDKN2A knockout HeLa whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 14 kDaLanes 1 - 2: Merged signal (red and green). Green - ab185620 observed at 14 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab185620 was shown to specifically react with CDKN2A/p14ARF in wild-type HeLa cells as signal was lost in CDKN2A knockout cells. Wild-type and CDKN2A knockout samples were subjected to SDS-PAGE. Ab185620 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. -
![Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-258221-anti-cdkn2a-p14arf-antibody-epr17878-western-blot.jpg)
Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate transfected with empty vector (vector control)
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate transfected with CDKN2A/p14ARF with His-tag
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 14 kDa
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-258219-anti-cdkn2a-p14arf-antibody-epr17878-western-blot.jpg)
Western blot - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)All lanes : Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620) at 1/1000 dilution
Lane 1 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression level of p14 ARF is relatively low in most cell types and will be induced by oncogene expression. (PMID: 9694807; PMID: 9694806).
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![Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-258218-anti-cdkn2a-p14arf-antibody-epr17878-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF or empty vector, labeling CDKN2A/p14ARF with ab185620 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab185620 at 1/250 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-258217-anti-cdkn2a-p14arf-antibody-epr17878-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with CDKN2A/p14ARF-GFP labeling CDKN2A/p14ARF with ab185620 at 1/120 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti Rabbit IgG (PE) at 1/150 dilution was used as the secondary antibody.
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![Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-258214-anti-cdkn2a-p14arf-antibody-epr17878-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)CDKN2A/p14ARF was immunoprecipitated from 1mg of CDKN2A/p14ARF transfected HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab185620 at 1/80 dilution.
Western blot was performed from the immunoprecipitate using ab185620 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: CDKN2A/p14ARF transfected HeLa whole cell lysate, 10µg (Input).
Lane 2: ab185620 IP in CDKN2A/p14ARF transfected HeLa whole cell lysate.
Lane 3: Rabbit IgG,monoclonal[EPR25A] - Isotype Control (ab172730) instead of ab185620 in CDKN2A/p14ARF transfected HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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![Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)](https://www.abcam.com/ps/products/185/ab185620/Images/ab185620-7-benefits-of-recombinant-antibodies.png)
Anti-CDKN2A/p14ARF antibody [EPR17878] (ab185620)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab185620 has been referenced in 3 publications.
- Miao J et al. Klotho retards renal fibrosis through targeting mitochondrial dysfunction and cellular senescence in renal tubular cells. Physiol Rep 9:e14696 (2021). PubMed: 33463897
- Horvat A et al. Helicobacter pylori pathogen regulates p14ARF tumor suppressor and autophagy in gastric epithelial cells. Oncogene 37:5054-5065 (2018). PubMed: 29849123
- Coleman KE et al. SENP8 limits aberrant neddylation of NEDD8 pathway components to promote cullin-RING ubiquitin ligase function. Elife 6:N/A (2017). PubMed: 28475037