Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)
![Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)](https://www.abcam.com/ps/products/26/ab26696/Images/ab26696-482439-AntiCDKN2A-p19ARF-antibody-immunocytochemistry-mef1.jpg "Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)")
Key features and details
- Rat monoclonal [5-C3] to CDKN2A/p19ARF
- Suitable for: WB, IP, ICC/IF, IHC-FoFr
- Reacts with: Mouse
- Isotype: IgG2b
Related conjugates and formulations
Overview
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Product name
Anti-CDKN2A/p19ARF antibody [5-C3]
See all CDKN2A/p19ARF primary antibodies -
Description
Rat monoclonal [5-C3] to CDKN2A/p19ARF -
Host species
Rat -
Tested applications
Suitable for: WB, IP, ICC/IF, IHC-FoFrmore details -
Species reactivity
Reacts with: Mouse
Does not react with: Hamster, Human -
Immunogen
Synthetic peptide corresponding to Mouse CDKN2A/p19ARF conjugated to keyhole limpet haemocyanin.
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Positive control
- 3T3 cell lysate positive control. ICC/IF: MEF1 cells
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General notes
The 5-C3 monoclonal has been knockout validated in WB and ICC/IF using wild type and knockout mouse samples. The data are shown in the images section.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.50
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
5-C3 -
Myeloma
Y3/Ag1.2.3 -
Isotype
IgG2b -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab26696 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 |
|---|---|---|
| WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa).
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| IP |
Use at an assay dependent concentration.
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| ICC/IF |
1/1000.
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| IHC-FoFr |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 19 kDa (predicted molecular weight: 17 kDa). |
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IP
Use at an assay dependent concentration. |
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ICC/IF
1/1000. |
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IHC-FoFr
Use at an assay dependent concentration. |
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
Nuclear -
Database links
- Entrez Gene: 12578 Mouse
- SwissProt: Q64364 Mouse
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Alternative names
- ARF antibody
- CDKN2A antibody
- Cyclin dependent kinase inhibitor 2A antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)
ab26696 staining CDKN2A/p19ARF in MEF1 cells. The cells were fixed with 4% PFA (10 mins), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab26696 at 5µg/ml and ab190573, Rabbit monoclonal to alpha Tubulin (Alexa Fluor® 647), at 2µg/ml (shown in red). The secondary antibody (shown in green) was ab150165, Alexa Fluor® 488 Goat anti-Rat IgG (H+L) used at a 1/1000 dilution for 1h at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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![Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)](https://www.abcam.com/ps/products/26/ab26696/Images/ab26696-37662-anti-cdkn2a-p19arf-antibody-5-c3-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)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). -
![Western blot - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)](https://www.abcam.com/ps/products/26/ab26696/Images/ab26696-482446-anti-cdkn2a-antibody-western-blot.jpg)
Western blot - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696) at 2 µg/ml + MEF-1 whole cell lysate at 20 µg
Secondary
Goat polyclonal to Rat IgG - H&L - Pre-Adsorbed (HRP) at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 19,85 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking buffer: 2% BSA
Gel type: MES
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![Western blot - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)](https://www.abcam.com/ps/products/26/ab26696/Images/ab26696-37664-anti-cdkn2a-p19arf-antibody-5-c3-western-blot.jpg)
Western blot - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)All lanes : Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)
Lane 1 : Total cell lysate from NIH3T3 cells with Arf gene deleted
Lane 2 : MEF total cell lysate
Secondary
All lanes : anti-rat IgG-HRP
Predicted band size: 17 kDa
Observed band size: 19 kDa why is the actual band size different from the predicted? -
![Immunoprecipitation - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)](https://www.abcam.com/ps/products/26/ab26696/Images/ab26696-37663-anti-cdkn2a-p19arf-antibody-5-c3-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CDKN2A/p19ARF antibody [5-C3] (ab26696)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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (20)
ab26696 has been referenced in 20 publications.
- Sun L et al. Nicotine triggers islet ß cell senescence to facilitate the progression of type 2 diabetes. Toxicology 441:152502 (2020). PubMed: 32473187
- Strauss RP et al. Loss of ARF/INK4A Promotes Liver Progenitor Cell Transformation Toward Tumorigenicity Supporting Their Role in Hepatocarcinogenesis. Gene Expr 20:39-52 (2020). PubMed: 32317048
- Chu B et al. ALOX12 is required for p53-mediated tumour suppression through a distinct ferroptosis pathway. Nat Cell Biol 21:579-591 (2019). PubMed: 30962574
- Lázaro S et al. Ablating all three retinoblastoma family members in mouse lung leads to neuroendocrine tumor formation. Oncotarget 8:4373-4386 (2017). PubMed: 27966456
- Lee CK et al. Syk-mediated tyrosine phosphorylation of mule promotes TNF-induced JNK activation and cell death. Oncogene 35:1988-95 (2016). PubMed: 26212014