Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18021] to p21 (Alexa Fluor® 647)
- Suitable for: ICC/IF, Flow Cyt
- Reacts with: Mouse
- Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm
Product nameAnti-p21 antibody [EPR18021] (Alexa Fluor® 647)
See all p21 primary antibodies
DescriptionRabbit monoclonal [EPR18021] to p21 (Alexa Fluor® 647)
ConjugationAlexa Fluor® 647. Ex: 652nm, Em: 668nm
SpecificityExpression levels of the target protein vary between different tissue/cell lines and in some cases induction may be required before a signal is observed.
Tested applicationsSuitable for: ICC/IF, Flow Cytmore details
Species reactivityReacts with: Mouse
Recombinant full length protein within Mouse p21 aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P39689
- ICC/IF: NIH/3T3 cells. Flow Cyt: NIH/3T3 cells.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or email@example.com.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab237265 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
This product gave a positive signal in NIH3T3 fixed with 4% formaldehyde (10 min).
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
- CDK-interacting protein 1 antibody
- CDKI antibody
Overlay histogram showing NIH/3T3 cells stained with ab237265 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS/10% normal Goat serum to block non-specific protein-protein interactions followed by the antibody (ab237265) (1x106 in 100 µl at 0.2 µg/ml (1/2500 dilution)) for 30 min at 22°C.
Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 647 (ab199093) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.
This antibody gave a positive signal in NIH/3T3 cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
ab237265 staining p21 in NIH/3T3 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab237265 at 1/1000 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab237265 has not yet been referenced specifically in any publications.