Anti-p21 antibody [CIP1/823] (ab220206)
Key features and details
- Mouse monoclonal [CIP1/823] to p21
- Suitable for: WB, ICC, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-p21 antibody [CIP1/823]
See all p21 primary antibodies -
Description
Mouse monoclonal [CIP1/823] to p21 -
Host species
Mouse -
Tested applications
Suitable for: WB, ICC, IHC-Pmore details -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant full length protein corresponding to Human p21 aa 1 to the C-terminus.
Database link: P38936 -
Positive control
- WB: HeLa cell lysate. ICC: HeLa cells. IHC-P: Human lung squamous cell carcinoma and bladder carcinoma tissues.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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: 0.05% Sodium azide
Constituents: 0.05% BSA, 99% PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
ab220206 was purified from Bioreactor Concentrate by Protein A/G. -
Clonality
Monoclonal -
Clone number
CIP1/823 -
Isotype
IgG2a -
Light chain type
kappa -
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 ab220206 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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WB |
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 18 kDa.
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ICC |
Use a concentration of 1 - 2 µg/ml.
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IHC-P |
Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
(Primary incubation for 30 minutes at RT). |
Notes |
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WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 18 kDa. |
ICC
Use a concentration of 1 - 2 µg/ml. |
IHC-P
Use a concentration of 2 - 4 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. (Primary incubation for 30 minutes at RT). |
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: 1026 Human
- Omim: 116899 Human
- SwissProt: P38936 Human
- Unigene: 370771 Human
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Alternative names
- CAP20 antibody
- CDK-interacting protein 1 antibody
- CDKI antibody
see all
Images
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All lanes : Anti-p21 antibody [CIP1/823] (ab220206) at 1 µg/ml
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : CDKN1A knockout HCT 116 cell lysate
Lane 3 : Wild-type MCF7 cell lysate
Lane 4 : CDKN1A knockout MCF7 cell lysate
Lane 5 : Wild-type A549 cell lysate
Lane 6 : CDKN1A knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 18 kDa
Observed band size: 21 kDa why is the actual band size different from the predicted?Western blot: Anti-CDKN1A antibody [CIP1/823] (ab220206) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab220206 was shown to bind specifically to CDKN1A. A band was observed at 21 kDa in wild-type cell lysates with no signal observed at this size in CDKN1A knockout cell lines. To generate this image, wild-type and CDKN1A knockout cell lysates were analysed. CDKN1A knockout cell lines in HCT 116 (ab288187), MCF7 (ab288200) and A549 (ab288213) backgrounds were used. 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
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Anti-p21 antibody [CIP1/823] (ab220206) at 1 µg/ml + HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Predicted band size: 18 kDaWestern blot analysis of HeLa cell lysate labeling p21 with ab220206 at 1 ug/ml.
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Immunocytochemistry analysis of HeLa cells labeling p21 with ab220206 at 1 ug/ml followed by Goat anti-Mouse IgG (Red). Membrane stained with Phalloidin 488 (Green).
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human bladder carcinoma tissue labeling p21 with ab220206 at 4 µg/ml.
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human lung squamous cell carcinoma tissue labeling p21 with ab220206 at 4 µg/ml.
Datasheets and documents
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Datasheet download
References (0)
ab220206 has not yet been referenced specifically in any publications.