Overview
- Product nameAnti-p21 antibodySee all p21 primary antibodies ...
- DescriptionRabbit polyclonal to p21
- SpecificityDetects a clean band at 21kDa corresponding to p21, as well as a cross-reacting band at 26kDa.
- Tested applicationsICC/IF, Sandwich ELISA, WB, IHC-P more details
- Species reactivityReacts with: Rat, Cow, Human
Predicted to work with: Cat, Dog - Immunogen
Synthetic peptide conjugated to KLH derived from within residues 100 to the C-terminus of Human p21.
(Peptide available as ab18623.)
- Positive controlThis antibody gave a positive signal in the following whole cell lysates: HeLa; Hela - Bleomycin Treated (40U/ml); HeLa Whole - Irradiated (5Gy); Hela - Hydroxyurea Treated (48hr, 1uM); PC12.
Properties
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
Our Abpromise guarantee covers the use of ab18209 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Notes |
|---|---|
| ICC/IF | |
| Sandwich ELISA | |
| WB | |
| IHC-P |
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Target
- 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. - 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.
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Database links
- Entrez Gene: 513497 Cow
- Entrez Gene: 474890 Dog
- Entrez Gene: 1026 Human
- Entrez Gene: 114851 Rat
- Omim: 116899 Human
- SwissProt: P38936 Human
- Unigene: 370771 Human
- Unigene: 10089 Rat
Target information above from: UniProt accession
P38936
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
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Alternative names
- Activating Fragment 1 antibodyActivating Fragment 1 antibodyCAP20 antibody
- CAP20 antibodyCation chloride cotransporter-interacting protein 1 antibodyCDK Interacting Protein 1 antibodyCDK Interacting Protein 1 antibodyCDK-interacting protein 1 antibodyCDKI antibodyCDKN 1 antibodyCDKN1 antibodyCDKN1 antibodyCDKN1A antibodyCDKN1A antibodyCDN1A_HUMAN antibodyCIP1 antibodyCIP1 antibodyCyclin Dependent Kinase Inhibitor 1A antibodyCyclin Dependent Kinase Inhibitor 1A antibodyCyclin-dependent kinase inhibitor 1 antibodyCyclin-dependent kinase inhibitor 1A (P21) antibodyCyclin-dependent kinase inhibitor 1A (p21, Cip1) antibodyDNA Synthesis Inhibitor antibodyMDA 6 antibodyMDA-6 antibodyMDA6 antibodyMDA6 antibodyMelanoma Differentiation Associated Protein 6 antibodyMelanoma differentiation-associated protein 6 antibodyMelanoma differentiation-associated protein antibodyp21 antibodyp21 antibodyP21 protein antibodyp21CIP1 antibodyp21Cip1/Waf1 antibodyp21WAF antibodyPIC1 antibodyPIC1 antibodySDI1 antibodySDI1 antibodySLC12A9 antibodyWAF1 antibodyWAF1 antibodyWild type p53 activated fragment 1 antibodyWildtype p53 Activating Fragment 1 antibodyWildtype p53 Activating Fragment 1 antibodyWildtype p53-activated fragment 1 antibody
see all
Anti-p21 antibody images
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Immunocytochemistry/ Immunofluorescence - p21 antibody (ab18209)ICC/IF image of ab18209 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab18209, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Immunocytochemistry/ Immunofluorescence - p21 antibody (ab18209)Ludovic Vallier, University of CambridgeThe image shows staining of human embryonic stem cells with ab18209. Staining appears to be nuclear. -
Western blot - p21 antibody (ab18209)Anti-p21 antibody (ab18209) at 1 µg/ml + HeLa whole cell lysate at 20 µg/ml
Secondary
Alexa Fluor Goat polyclonal to Rabbit IgG (700) at 1/10000 dilution
Predicted band size : 21 kDa
Observed band size : 21 kDa
Additional bands at : 26 kDa (possible cross reactivity). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - p21 antibody (ab18209)IHC image of p21 staining in human liver carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18209, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. -
Western blot - p21 antibody (ab18209)All lanes : Anti-p21 antibody (ab18209) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Hela Whole Cell Lysate - Bleomycin Treated (40U/ml)
Lane 3 : HeLa Whole Cell Lysate - Irradiated (5Gy)
Lane 4 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 1uM)
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 21 kDa
Observed band size : 21 kDa
Additional bands at : 36 kDa,56 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 20 minutes -
Sandwich ELISA - p21 antibody (ab18209)Standard curve for p21 (Analyte: ab56278); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [AC8.3] to p21 (ab118) at 5µg/ml and Detector Antibody Rabbit polyclonal to p21 (ab18209) at 0.5µg/ml. -
Western blot - p21 antibody (ab18209)Anti-p21 antibody (ab18209) at 1 µg/ml + PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 21 kDa
Observed band size : 21 kDa
Additional bands at : 40 kDa,80 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 20 minutes -
Western blot - Anti-p21 antibody (ab18209)Anti-p21 antibody (ab18209) at 1 µg/ml +p21 protein (Tagged) (ab56278) at 0.01 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 150 seconds
Protocols
References for Anti-p21 antibody (ab18209)
This product has been referenced in:
- Symanowski J et al. A histone deacetylase inhibitor LBH589 downregulates XIAP in mesothelioma cell lines which is likely responsible for increased apoptosis with TRAIL. J Thorac Oncol 4:149-60 (2009). Read more (PubMed: 19179889) »
