Recombinant Anti-CDKN2A/p16INK4a 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...
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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 |
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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. |
IP
1/30. |
IHC-P
1/500. |
WB
1/1000. Predicted molecular weight: 17 kDa. |
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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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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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)
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)
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)
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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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 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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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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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
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