Anti-p21 (phospho T145) antibody (ab47300)
Key features and details
- Rabbit polyclonal to p21 (phospho T145)
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-p21 (phospho T145) antibody
See all p21 primary antibodies -
Description
Rabbit polyclonal to p21 (phospho T145) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human p21 aa 100-200 (phospho T145).
Database link: P38936 -
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.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
The antibody was purified using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site. -
Primary antibody notes
p21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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.
|
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| WB |
1/500 - 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 18 kDa).
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| IHC-P | (1) |
Use at an assay dependent concentration.
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| Notes |
|---|
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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). |
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IHC-P
Use at an assay dependent concentration. |
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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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 -
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. -
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.
Protocols
Datasheets and documents
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Datasheet download
References (10)
ab47300 has been referenced in 10 publications.
- Feng C et al. Tumor-Suppressive MicroRNA-216b Binds to TPX2, Activating the p53 Signaling in Human Cutaneous Squamous Cell Carcinoma. Mol Ther Nucleic Acids 20:186-195 (2020). PubMed: 32169806
- Chen YS et al. MiR-1908/EXO1 and MiR-203a/FOS, regulated by scd1, are associated with fracture risk and bone health in postmenopausal diabetic women. Aging (Albany NY) 12:9549-9584 (2020). PubMed: 32454462
- Chen W et al. GATA6 Exerts Potent Lung Cancer Suppressive Function by Inducing Cell Senescence. Front Oncol 10:824 (2020). PubMed: 32596145
- Stafman LL et al. Targeting PIM kinase as a therapeutic strategy in human hepatoblastoma. Oncotarget 9:22665-22679 (2018). PubMed: 29854306
- Yang J et al. Combination of IFITM1 knockdown and radiotherapy inhibits the growth of oral cancer. Cancer Sci 109:3115-3128 (2018). PubMed: 29770536
- Song J et al. Melatonin induces the apoptosis and inhibits the proliferation of human gastric cancer cells via blockade of the AKT/MDM2 pathway. Oncol Rep 39:1975-1983 (2018). PubMed: 29484412
- Kawano M et al. MicroRNA-20b promotes cell proliferation via targeting of TGF-ß receptor II and upregulates MYC expression in Ewing's sarcoma cells. Int J Oncol 51:1842-1850 (2017). PubMed: 29039480
- 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 . PubMed: 26243299
- Liu R et al. Itraconazole suppresses the growth of glioblastoma through induction of autophagy: involvement of abnormal cholesterol trafficking. Autophagy 10:1241-55 (2014). PubMed: 24905460
- Pitre A et al. Synemin promotes AKT-dependent glioblastoma cell proliferation by antagonizing PP2A. Mol Biol Cell 23:1243-53 (2012). WB ; Human . PubMed: 22337773
