Name: Anti-CDKN2A/p16INK4a antibody [EPR24167-43]
SKU: ab270058
Rating: 4 (1 reviews)

Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) is a rabbit monoclonal antibody that is used to detect CDKN2A/p16INK4a in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human samples.

- Specificity confirmed with CDKN2A/p16INK4a knockout cell line validation

Images

Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (AB270058), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59.94% 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

	ICC/IF	IP	WB	IHC-P	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Recombinant full length protein - Human	Not recommended	Not recommended	Tested	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Recombinant full length protein - Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Recombinant full length protein - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Recombinant full length protein - Human	Dilution info -	Notes -

Associated Products

Select an associated product type

1 product for Alternative Version

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: 59.94% 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: EPR24167-43
Purification technique: Affinity purification Protein A
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

What is this antibody validated in?

Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human samples.

What is the molecular weight of CDKN2A/p16INK4a?

Anti-CDKN2A/p16INK4a [EPR24167-43] (ab270058) specifically detects a band for CDKN2A/p16INK4a (UniProt: P42771) at a molecular weight of 17kDa.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) has been confirmed by Western blot testing in CDKN2A Knockout HEK293T cells.

Other related products

We have a range of other formats of antibody clone [EPR24167-43] also available for your convenience:
ab270058, Carrier free - ab282285

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.

11 product images

Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
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.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 2: HEK-293 (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 3: Saos-2 (human osteosarcoma epithelial), whole cell lysate at 20 µg
Lane 4: MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 5: MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16 kDa
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
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.
All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058) at 1/1000 dilution
Lane 1: His-tagged human CDKN2A recombinant protein at 0.01 µg
Lane 2: GST-tagged human CDKN2B recombinant protein 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 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a 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.
Immunoprecipitation - Anti-CDKN2A/p16INK4a 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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab270058 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
All lanes: Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
Predicted band size: 17 kDa
Observed band size: 16 kDa
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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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.
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.
Immunoprecipitation - Anti-CDKN2A/p16INK4a 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)(VeriBlot for IP Detection Reagent (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab270058 in HEK-293 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 32 seconds
All lanes: Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
Predicted band size: 17 kDa
Observed band size: 16 kDa
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: MCF7 (PMID: 10914724).
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
Western blot: Anti-CDKN2A antibody [EPR24167-43] (ab270058) staining at 1/2000 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, ab270058 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 [EPR24167-43] (ab270058) at 1/2000 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
Western blot - Anti-CDKN2A/p16INK4a antibody [EPR24167-43] (ab270058)
Western blot: Anti-CDKN2A antibody [EPR24167-43] (ab270058) staining at 1/1000 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, ab270058 was shown to bind specifically to CDKN2A. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% 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 [EPR24167-43] (ab270058) at 1/1000 dilution
Lane 1: HeLa cell lysate at 20 µg
Lane 2: HEK-293 cell lysate at 20 µg
Lane 3: Saos-2 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 17 kDa