Key features and details
- Rabbit polyclonal to p21 (phospho T145)
- Suitable for: ICC/IF, WB, IHC-P, ELISA
- Reacts with: Human
- Isotype: IgG
Product nameAnti-p21 (phospho T145) antibody
See all p21 primary antibodies
DescriptionRabbit polyclonal to p21 (phospho T145)
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, ELISAmore details
Species reactivityReacts with: Human
synthesized phosphopeptide derived from human p21Cip1 around the phosphorylation site of threonine 145 (R-Q-TP-S-M)
- Human breast carcinoma tissue and EGF treated HeLa cell extracts
p21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 50% Glycerol, 0.87% Sodium chloride
Concentration information loading...
PurityImmunogen affinity purified
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.
Primary antibody notesp21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145.
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.
|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.|
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.
Tissue specificityExpressed in all adult human tissues, with 5-fold lower levels observed in the brain.
Sequence similaritiesBelongs to the CDI family.
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.
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
- CAP20 antibody
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All lanes : Anti-p21 (phospho T145) antibody (ab47300)
Lane 1 : EGF treated HeLa cells
Lane 2 : HeLa cells
Predicted band size: 18 kDa
ab47300 staining human breast carcinoma tissue by IHC-P (left hand panel). The right hand panel shows staining in the presence of phospho-peptide.
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.
ab47300 has been referenced in 7 publications.
- 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