Recombinant Anti-CDKN2A/p16INK4a antibody [EPR20418] (ab211542)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20418] to CDKN2A/p16INK4a
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CDKN2A/p16INK4a antibody [EPR20418]
See all CDKN2A/p16INK4a primary antibodies -
Description
Rabbit monoclonal [EPR20418] to CDKN2A/p16INK4a -
Host species
Rabbit -
Specificity
Expression levels of the CDKN2A/p16INK4a protein may vary with sample type. It's 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.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse
Does not react with: Rat, Human -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: MEF whole cell lysate; His-tagged mouse CDKN2A/p16INK4a recombinant protein, aa1-168. ICC/IF: MEF cells. Flow Cyt (intra): MEF cells. IP: MEF whole cell lysate.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol, PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20418 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab211542 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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Flow Cyt (Intra) |
1/500.
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WB | (2) |
1/2000. Detects a band of approximately 16, 14 kDa (predicted molecular weight: 18 kDa).
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IP |
1/30.
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ICC/IF | (2) |
1/100.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/2000. Detects a band of approximately 16, 14 kDa (predicted molecular weight: 18 kDa). |
IP
1/30. |
ICC/IF
1/100. |
Target
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Cellular localization
Cytoplasmic and Nuclear -
Database links
- Entrez Gene: 12578 Mouse
- SwissProt: P51480 Mouse
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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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Immunofluorescent analysis of 100% methanol-fixed MEF (mouse embryonic fibroblast cell line) and NIH/3T3 (mouse embyro fibroblast cell line) cells labeling CDKN2A/p16INK4a with ab211542 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution (green). Confocal image showing nuclear and cytoplasmic staining on MEF cell line.
Negative control: NIH/3T3 (PMID: 15210712).
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1,000 dilution.
MEF cells were kindly provided by professor Pinlong Xu, Zhejiang University.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed MEF (mouse embryonic fibroblast cell line) cell line labeling CDKN2A/p16INK4awith ab211542 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
MEF cells were kindly provided by professor Pinlong Xu, Zhejiang University.
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All lanes : Anti-CDKN2A/p16INK4a antibody [EPR20418] (ab211542) at 1/1000 dilution
Lane 1 : His-tagged mouse CDKN2A/p16INK4a recombinant protein
Lane 2 : His-tagged mouse CDKN2B/p15INK4b 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
Developed using the ECL technique.
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-CDKN2A/p16INK4a antibody [EPR20418] (ab211542) at 1/2000 dilution
Lane 1 : MEF (mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
Lane 3 : Mouse lung lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Mouse testis lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 18 kDa
Observed band size: 14,16 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
Negative control: NIH/3T3 (PMID: 15210712).
Limited expression in mouse normal tissues has been documented in the literature (PMID: 9244355).
14 kDa band is a proteolytic p16INK4a lacking C-terminus (PMID: 18053084).
MEF cells are kindly provided by professor Pinlong Xu, Zhejiang University.
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CDKN2A/p16INK4a was immunoprecipitated from fresh MEF lysates with ab211542 at 1/30 dilution (2ug in 0.35mg lysates). Lysates were made fresh and used immediately to minimize protein degradation. Western blot was performed on the immunoprecipitate using ab211542 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: MEF (mouse embryo fibroblast) whole cell lysate 10 μg (IP lysis buffer)
Lane 2: MEF (mouse embryo fibroblast) whole cell lysate (IP lysis buffer)
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211542 in MEF whole cell lysate (IP lysis buffer)Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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CDKN2A/p16INK4a was immunoprecipitated from 0.35 mg of MEF (mouse embryonic fibroblast cell line) whole cell lysate with ab211542 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab211542 at 1/5000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: MEF whole cell lysate 10 μg (Input).
Lane 2: ab211542 IP in MEF whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211542 in MEF whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.
MEF cells were kindly provided by professor Pinlong Xu, Zhejiang University.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (53)
ab211542 has been referenced in 53 publications.
- Sun J et al. Growth differentiation factor 11 accelerates liver senescence through the inhibition of autophagy. Aging Cell 21:e13532 (2022). PubMed: 34905649
- Adili A et al. Atrial Fibrillation Underlies Cardiomyocyte Senescence and Contributes to Deleterious Atrial Remodeling during Disease Progression. Aging Dis 13:298-312 (2022). PubMed: 35111375
- Safwan-Zaiter H et al. Dynamic Spatiotemporal Expression Pattern of the Senescence-Associated Factor p16Ink4a in Development and Aging. Cells 11:N/A (2022). PubMed: 35159350
- Kim H et al. Attenuation of intrinsic ageing of the skin via elimination of senescent dermal fibroblasts with senolytic drugs. J Eur Acad Dermatol Venereol 36:1125-1135 (2022). PubMed: 35274377
- Limberger T et al. KMT2C methyltransferase domain regulated INK4A expression suppresses prostate cancer metastasis. Mol Cancer 21:89 (2022). PubMed: 35354467