- Datasheet
- Specific References (4)
- Protocols
Overview
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Product nameAnti-p21 (phospho T145) antibody
See all p21 primary antibodies -
DescriptionRabbit polyclonal to p21 (phospho T145)
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, WB, IHC-P, ELISAmore details
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Species reactivityReacts with: Human
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Immunogen
synthesized phosphopeptide derived from human p21Cip1 around the phosphorylation site of threonine 145 (R-Q-TP-S-M)
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Positive control
- Human breast carcinoma tissue and EGF treated HeLa cell extracts
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General notes
p21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
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Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesThe antibody was purified using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
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Primary antibody notesp21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
Applications
Our Abpromise guarantee covers the use of ab47300 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 |
|---|---|---|
| ICC/IF | Use a concentration of 1 µg/ml. | |
| WB | 1/500 - 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 18 kDa). | |
| IHC-P | Use at an assay dependent concentration. | |
| ELISA | 1/10000. |
Target
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FunctionMay 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.
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Tissue specificityExpressed in all adult human tissues, with 5-fold lower levels observed in the brain.
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Sequence similaritiesBelongs to the CDI family.
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DomainThe 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 localizationCytoplasm. 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 (phospho T145) antibody (ab47300)ab47300 staining human breast carcinoma tissue by IHC-P (left hand panel). The right hand panel shows staining in the presence of phospho-peptide. -
Western blot - Anti-p21 (phospho T145) antibody (ab47300)All lanes : Anti-p21 (phospho T145) antibody (ab47300)
Lane 1 : EGF treated HeLa cells
Lane 2 : HeLa cells
Predicted band size: 18 kDa -
Immunocytochemistry/ Immunofluorescence - Anti-p21 (phospho T145) antibody (ab47300)ICC/IF image of ab47300 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47300, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References
This product has been referenced in:
- Stafman LL et al. Targeting PIM kinase as a therapeutic strategy in human hepatoblastoma. Oncotarget 9:22665-22679 (2018). Read more (PubMed: 29854306) »
- Kawano M et al. microRNA-93 promotes cell proliferation via targeting of PTEN in Osteosarcoma cells. J Exp Clin Cancer Res 34:76 (2015). WB . Read more (PubMed: 26243299) »
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