Name: Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal
SKU: ab108349
Rating: 4 (7 reviews)

Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) is a rabbit monoclonal antibody detecting CDKN2A/p16INK4a in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P. Suitable for Human.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 190 publications

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (AB108349), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Not recommended	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes For unpurified use at 1/10 - 1/100. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes For unpurified use at 1/1000 - 1/10000.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes For unpurified use at 1/100 - 1/250.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/270 - 1/500	Notes For unpurified use at 1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes For unpurified use at 1/250 - 1/500. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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Target data

See full target information CDKN2A

Function

The protein expressed by the gene CDKN2A acts as a negative regulator of normal cell proliferation by strongly interacting with CDK4 and CDK6. This interaction inhibits the ability of CDK4 and CDK6 to interact with cyclins D and to phosphorylate the retinoblastoma protein. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

CDKN2, MTS1, CDKN2A, Cyclin-dependent kinase inhibitor 2A, Cyclin-dependent kinase 4 inhibitor A, Multiple tumor suppressor 1, p16-INK4a, CDK4I, MTS-1, p16-INK4, p16INK4A

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR1473
Purification technique: Affinity purification Protein A
Specificity: Expression levels of the CDKN2A/p16INK4a protein may vary with sample type. It is barely expressed in normal tissue, and mostly expressed in some tumour tissues, such as cervical cancer, breast cancer and so on. Moreover, only expressed in some cell lines. Please see images for recommended positive controls.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), IHC-P, IP and WB.

Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) was first used in a scientific publication in 2013 and has been cited over 192 times in peer reviewed journals. It's performance in Western Blot and IHC in human samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) has been confirmed by Western Blot testing in CDKN2A/p16INK4a knockout HEK-293T cells.

Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) has 6 independent reviews from customers.

Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) specifically detects CDKN2A/p16INK4a (UniProt ID: P42771; Molecular weight: 17kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Conjugation-ready, carrier free format available for antibody clone EPR1473 - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free ab186932.

Antibody clone EPR1473 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 647, PE (Alexa Fluor® 647 Anti-CDKN2A/p16INK4a antibody [EPR1473] ab192054, PE Anti-CDKN2A/p16INK4a antibody [EPR1473] ab209579).

CDKN2A, encoding the tumor suppressor protein p16INK4a, plays a crucial role in regulating the cell cycle by inhibiting cyclin-dependent kinases, thereby preventing uncontrolled cell proliferation. Mutations or deletions in CDKN2A are frequently associated with various cancers, including melanoma, pancreatic cancer and head and neck squamous cell carcinoma.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

15 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling CDKN2A/p16INK4a with unpurified ab108349 at a dilution of 1/250.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Image from Shan M et al., PLoS One. 2013;8(10):e76408. Fig 1.; doi: 10.1371/journal.pone.0076408. eCollection 2013. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Formalin-fixed, paraffin-embedded human normal breast, luminal-A DCIS (ductal carcinoma in situ) and triple negative breast cancer tissues stained for CDKN2A/p16INK4a using ab108349 in immunohistochemical analysis.
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
5% NFDM/TBST was used as a blocking and diluting buffer.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/1000 dilution
Lane 1: Human CDKN2A full-length recombinant protein with His-tag at 0.01 µg
Lane 2: Human CDKN2B full-length recombinant protein with GST-tag at 0.01 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDa
Exposure time: 10s
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/1000 dilution
Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
Lane 2: HEK-293T (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg
Lane 3: Saos-2 (Human osteosarcoma cell line) cell lysate at 10 µg
Predicted band size: 17 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling CDKN2A/p16INK4a with purified ab108349 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, an HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).
Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
ab108349 (purified) at 1/30 immunoprecipitating CDKN2A/p16INK4a in HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Predicted band size: 17 kDa
Observed band size: 17 kDa
Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Overlay histogram showing HEK-293 (Human epithelial cell line from embryonic kidney) cells stained with unpurified ab108349 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab108349, 1/100) for 30 minutes at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK-293 cells fixed with 80% methanol (5 minutes)/permeabilized with 0.1% PBS-Tween for 20 minutes used under the same conditions.
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/2000 dilution
Lane 1: HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg
Lane 2: Saos-2 (Human osteosarcoma cell line) cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDa
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/2000 dilution
All lanes: HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 17 kDa
Observed band size: 17 kDa
Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Intracellular Flow Cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling CDKN2A/p16INK4a with purified ab108349 at 1/270 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabeled control, cells without incubation with primary and secondary antibodies.
Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Flow cytometry overlay histogram showing left HeLa positive cells and right negative MCF7 stained with ab108349 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab108349) (1x 106 in 100μl at 0.04μg/ml (1/52500)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling CDKN2A/p16INK4a with ab108349 at a concentration of 1µg/ml. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.
ab108349 anti-CDKN2A/p16INK4a antibody [EPR1473] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Western blot: Anti-CDKN2A antibody [EPR1473] (ab108349) staining at 1/4000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108349 was shown to bind specifically to CDKN2A. A band was observed at 17 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in CDKN2A knockout cell line. To generate this image, wild-type and CDKN2A knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349) at 1/4000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: CDKN2A knockout HEK-293T cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 17 kDa
Observed band size: 17 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Tissue Microarrays stained for "Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal" using "ab108349"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab108349 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (ab108349)
Immunohistochemical analysis of formalin fixed paraffin embedded human testis labelling CDKN2A/p16INK4a with ab108349 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab108349 anti-CDKN2A/p16INK4a antibody [EPR1473] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)