Recombinant Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
![Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-465175-anti-cdkn2ap16ink4a-antibody-epr2416743-western-blot-adenocarcinoma.jpg "Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR24167-43] to CDKN2A/p16INK4a
- Suitable for: ICC/IF, IP, IHC-P, WB, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CDKN2A/p16INK4a antibody [EPR24167-43]
See all CDKN2A/p16INK4a primary antibodies -
Description
Rabbit monoclonal [EPR24167-43] to CDKN2A/p16INK4a -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, IHC-P, WB, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, HEK-293, Saos-2, His-tagged human CDKN2A recombinant protein lysates, human CDKN2A full-length recombinant protein with His-tag, human CDKN2B full-length recombinant protein with GST-tag. IHC-P: Human cervical cancer and human endometrial cancer tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa, HEK-293 cells.
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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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR24167-43 -
Isotype
IgG -
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 ab270058 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 |
|---|---|---|
| ICC/IF |
1/50.
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| IP |
1/30.
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| IHC-P | (1) |
1/500.
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| WB |
1/1000. Predicted molecular weight: 17 kDa.
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| Flow Cyt (Intra) |
1/500.
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| Notes |
|---|
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ICC/IF
1/50. |
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IP
1/30. |
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IHC-P
1/500. |
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WB
1/1000. Predicted molecular weight: 17 kDa. |
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Flow Cyt (Intra)
1/500. |
Target
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Cellular localization
Cytoplasmic and Nuclear -
Database links
- Entrez Gene: 1029 Human
- Omim: 600160 Human
- SwissProt: P42771 Human
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Form
There are 4 isoforms produced by alternative splicing. Isoform 1 also known as: p16INK4a; Isoform 3 also known as: p12; Isoform 4 also known as: p14ARF; p19ARF; ARF. -
Alternative names
- CCM2 antibody
- CDK4 inhibitor p16 INK4 antibody
- CDK4I antibody
see all
Images
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Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)All lanes : Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
Lane 3 : Saos-2 (human osteosarcoma epithelial), whole cell lysate
Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
Lane 5 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 17 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: MCF7, MDA-MB-231 (PMID: 10914724)
The MW observed is consistent with the literature. (PMID: 18053084)
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 70 seconds.
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![Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-465176-anti-cdkn2ap16ink4a-antibody-epr2416743-western-blot-recombinant.jpg)
Western blot - Anti-CDKN2A antibody [EPR24167-43] (ab270058)All lanes : Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution
Lane 1 : His-tagged human CDKN2A recombinant protein
Lane 2 : GST-tagged human CDKN2B recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 17 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
This antibody specifically recognizes CDKN2A recombinant protein, but not CDKN2B recombinant protein.
Exposure time: 3.25 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-1-anti-cdkn2ap16ink4a-antibody-epr2416743-immunohistochemistry-human-cervical-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human cervical cancer (PMID: 9736404). The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-2-anti-cdkn2ap16ink4a-antibody-epr2416743-immunohistochemistry-human-endometrial-cancer.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining in human endometrial cancer. The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-3-anti-cdkn2ap16ink4a-antibody-epr2416743-immunohistochemistry-human-cervix.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A antibody [EPR24167-43] (ab270058)Immunohistochemical analysis of paraffin-embedded Human cervix tissue labelling CDKN2A/p16INK4a with ab270058 at 1/500 (0.936 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in human cervix (PMID: 9736404). The section was incubated with ab270058 for 30 minutes at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 minutes.
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![Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-4-anti-cdkn2ap16ink4a-antibody-epr2416743-immunocytochemistry-hela.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling CDKN2A/p16INK4a with ab270058 at 1/50 (9.36 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear and cytoplasmic staining in HeLa cell line. Negative control: MCF7 (PMID: 10914724) is observed.
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-5-anti-cdkn2ap16ink4a-antibody-epr2416743-flow-cytometry-mcf7.jpg)
Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 ( Human breast adenocarcinoma epithelial cell, Left)/ HeLa (Human cervix adenocarcinoma epithelial cell, Right) cells labelling CDKN2A/p16INK4a with ab270058 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: MCF7 (PMID: 10914724).
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![Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-6-anti-cdkn2ap16ink4a-antibody-epr2416743-immunoprecipitation-hela.jpg)
Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab270058 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270058 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug
Lane 2 : ab270058 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270058 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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![Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-7-anti-cdkn2ap16ink4a-antibody-epr2416743-immunoprecipitation-hek293.jpg)
Immunoprecipitation - Anti-CDKN2A antibody [EPR24167-43] (ab270058)CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell), whole cell lysate 10 ug with ab270058 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270058 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: HEK-293 (human embryonic kidney epithelial cell), whole cell lysate 10 ug
Lane 2: ab270058 IP in HEK-293 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab270058 in HEK-293 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
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![Anti-CDKN2A antibody [EPR24167-43] (ab270058)](https://www.abcam.com/ps/products/270/ab270058/Images/ab270058-8-Benefits-of-Recombinant-antibodies.jpg)
Anti-CDKN2A antibody [EPR24167-43] (ab270058)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (4)
ab270058 has been referenced in 4 publications.
- Ye Z et al. LMCD1 facilitates the induction of pluripotency via cell proliferation, metabolism, and epithelial-mesenchymal transition. Cell Biol Int 46:1409-1422 (2022). PubMed: 35842772
- Yan L et al. A ferroptosis associated gene signature for predicting prognosis and immune responses in patients with colorectal carcinoma. Front Genet 13:971364 (2022). PubMed: 36160009
- Ji ML et al. Sirt6 attenuates chondrocyte senescence and osteoarthritis progression. Nat Commun 13:7658 (2022). PubMed: 36496445
- Fang F et al. m6A RNA methylation regulator-based signature for prognostic prediction and its potential immunological role in uterine corpus endometrial carcinoma. BMC Cancer 22:1364 (2022). PubMed: 36581816