Skip to main content

Knockout Tested Rabbit Recombinant Monoclonal p16INK4A antibody. Carrier free. Suitable for IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 3 publications.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (AB186932), expandable thumbnail
  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (AB186932), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (AB186932), expandable thumbnail
  • Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (AB186932), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (AB186932), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form
Liquid
Clonality
Monoclonal

Immunogen

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

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFIPWBFlow Cyt (Intra)IHC-P
Human
Not recommended
Tested
Tested
Tested
Tested

Not recommended
Not recommended

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species
Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
-
Notes

Please check the parent abID, Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349, for a recommended dilution.

Tested
Tested

Species
Human
Dilution info
-
Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species
Human
Dilution info
-
Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

12 products for Alternative Product

2 products for Alternative Version

Target data

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

Recommended products

Knockout Tested Rabbit Recombinant Monoclonal p16INK4A antibody. Carrier free. Suitable for IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 3 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free
Yes
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
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

ab186932 is the carrier-free version of Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349.

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

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.

10 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail
    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] - BSA and Azide free (ab186932)

    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 Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349 in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

    All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    All lanes: HEK293 (human embryonic kidney) whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051)

    Predicted band size: 17 kDa

    Exposure time: 3min

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labeling CDKN2A/p16INK4a with purified Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

  • Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

    All lanes: Immunoprecipitation - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (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] - BSA and Azide free (ab186932), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Overlay histogram showing HEK-293 (Human epithelial cell line from embryonic kidney) cells stained with unpurified Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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 (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349, 1/100) for 30 minutes at 22ºC. 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 22ºC. 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labeling CDKN2A/p16INK4a with unpurified Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349 at a dilution of 1/250.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Flow Cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling CDKN2A/p16INK4a with purified Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

  • Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Western blot - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    This data was developed using the same antibody clone in a different buffer formulation (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).

    Western blot: Anti-CDKN2A antibody [EPR1473] (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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, Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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 (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] - BSA and Azide free (ab186932), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    Tissue Microarrays stained for "Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal" using "Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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 Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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.

  • Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - BSA and Azide free (ab186932)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal ab108349).
    Flow cytometry overlay histogram showing left HeLa positive cells and right negative MCF7 stained with Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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 (Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal 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.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com