Recombinant Anti-p21 antibody [EPR18021] (ab188224)
![Western blot - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279596-anti-p21-antibody-epr18021-western-blot.jpg "Western blot - Anti-p21 antibody [EPR18021] (ab188224)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18021] to p21
- Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-p21 antibody [EPR18021]
See all p21 primary antibodies -
Description
Rabbit monoclonal [EPR18021] to p21 -
Host species
Rabbit -
Specificity
Expression levels of the target protein vary between different tissue/cell lines and in some cases induction may be required before a signal is observed. -
Tested applications
Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: RAW 264.7 whole cell lysate; NIH/3T3, untreated and treated with 1µM staurosporine for 2hrs, whole cell lysates. IHC-P: Mouse testis and lung tissues. ICC/IF: RAW 264.7 and NIH/3T3 cells. Flow Cyt (intra): RAW 264.7 cells. IP: NIN/3T3 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: 59% PBS, 0.05% BSA, 40% Glycerol -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18021 -
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 ab188224 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 |
|---|---|---|
| WB | (1) |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
|
| IHC-P | (4) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
| IP |
1/30.
|
|
| ICC/IF | (2) |
1/500.
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| Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa). |
|
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
IP
1/30. |
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ICC/IF
1/500. |
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
May be the important intermediate by which p53/TP53 mediates its role as an inhibitor of cellular proliferation in response to DNA damage. Binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. Functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, inhibits the kinase activity of the cyclin D-CDK4 complex. -
Tissue specificity
Expressed in all adult human tissues, with 5-fold lower levels observed in the brain. -
Sequence similarities
Belongs to the CDI family. -
Domain
The PIP-box K+4 motif mediates both the interaction with PCNA and the recuitment of the DCX(DTL) complex: while the PIP-box interacts with PCNA, the presence of the K+4 submotif, recruits the DCX(DTL) complex, leading to its ubiquitination.
The C-terminal is required for nuclear localization of the cyclin D-CDK4 complex. -
Post-translational
modificationsPhosphorylation of Thr-145 by Akt or of Ser-146 by PKC impairs binding to PCNA. Phosphorylation at Ser-114 by GSK3-beta enhances ubiquitination by the DCX(DTL) complex.
Ubiquitinated by MKRN1; leading to polyubiquitination and 26S proteasome-dependent degradation. Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, leading to its degradation during S phase or following UV irradiation. Ubiquitination by the DCX(DTL) complex is essential to control replication licensing and is PCNA-dependent: interacts with PCNA via its PIP-box, while the presence of the containing the 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to its degradation. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 12575 Mouse
- SwissProt: P39689 Mouse
- Unigene: 195663 Mouse
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Alternative names
- CAP20 antibody
- CDK-interacting protein 1 antibody
- CDKI antibody
see all
Images
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Western blot - Anti-p21 antibody [EPR18021] (ab188224)All lanes : Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution
Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 1µM staurosporine for 2hrs, 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
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The expression level of p21 protein can be induced using staurosporine (protein kinase C inhibitor) (PMID:7677742).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279594-anti-p21-antibody-epr18021-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on mouse testis is observed (PMID: 9170103).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279588-anti-p21-antibody-epr18021-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on RAW264.7 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279585-anti-p21-antibody-epr18021-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-p21 antibody [EPR18021] (ab188224)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21with ab188224 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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![Western blot - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279597-anti-p21-antibody-epr18021-western-blot.jpg)
Western blot - Anti-p21 antibody [EPR18021] (ab188224)Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution + RAW264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 8 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-11-anti-p21-antibody-epr18021-westernblot-raw-264-7-mouse.jpg)
Western blot - Anti-p21 antibody [EPR18021] (ab188224)Different batches of ab188224 were tested on RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 18 kDa. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279593-anti-p21-antibody-epr18021-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Sporadic nuclear staining on mouse lung is observed (PMID: 25333671).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279587-anti-p21-antibody-epr18021-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on NIH/3T3 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Immunoprecipitation - Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-279583-anti-p21-antibody-epr18021-immunoprecipitation.jpg)
Immunoprecipitation - Anti-p21 antibody [EPR18021] (ab188224)p21 was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab188224 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab188224 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: NIH/3T3 whole cell lysate 10 µg (Input).
Lane 2: ab188224 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188224 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
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![Anti-p21 antibody [EPR18021] (ab188224)](https://www.abcam.com/ps/products/188/ab188224/Images/ab188224-10-benefits-of-recombinant-antibodies.png)
Anti-p21 antibody [EPR18021] (ab188224)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (87)
ab188224 has been referenced in 87 publications.
- Shaalan AK et al. Inhibition of Aurora Kinase B activity disrupts development and differentiation of salivary glands. Cell Death Discov 7:16 (2021). PubMed: 33462217
- Buitrago-Molina LE et al. p53-Independent Induction of p21 Fails to Control Regeneration and Hepatocarcinogenesis in a Murine Liver Injury Model. Cell Mol Gastroenterol Hepatol 11:1387-1404 (2021). PubMed: 33484913
- Cheng X et al. Long non-coding RNA Meg3 deficiency impairs glucose homeostasis and insulin signaling by inducing cellular senescence of hepatic endothelium in obesity. Redox Biol 40:101863 (2021). PubMed: 33508742
- Bernasocchi T et al. Dual functions of SPOP and ERG dictate androgen therapy responses in prostate cancer. Nat Commun 12:734 (2021). PubMed: 33531470
- Tashjian RZ et al. The effect of estrogen-like compound on rotator cuff tendon healing in a murine model. J Orthop Res N/A:N/A (2021). PubMed: 33533088