Overview
Product name
Human p21 ELISA KitSee all p21 kitsDetection method
ColorimetricPrecision
Intra-assay Sample n Mean SD CV% MCF7 lysates 5 4.2% Inter-assay Sample n Mean SD CV% MCF7 lysates 3 7.3% Sample type
Cell culture extracts, Tissue LysateAssay type
Sandwich (quantitative)Sensitivity
11 pg/mlRange
62.5 pg/ml -4000 pg/mlRecovery
Sample specific recovery Sample type Average % Range Cell culture media 120 115%- 129% Assay time
1h30mAssay duration
One step assaySpecies reactivity
Reacts with:HumanProduct overview
Human p21 ELISA Kit (ab214658) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of p21 protein in tissue lysate and cell culture extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human p21 with 11 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less- High sensitivity, specificity and reproducibility from superior antibodies- Fully validated in biological samples- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
Notes
p21 plays a critical role in the cellular response to DNA damage, and its overexpression results in cell cycle arrest. Upregulation of p21 mRNA and protein following ionizing radiation is dependent on p53 (TP53), and p21 mediates cell cycle arrest in response to the p53 checkpoint pathway. p21 binds to and inhibits cyclin-dependent kinase activity, preventing phosphorylation of critical cyclin-dependent kinase substrates and blocking cell cycle progression. p21 functions in the nuclear localization and assembly of cyclin D-CDK4 complex and promotes its kinase activity towards RB1. At higher stoichiometric ratios, p21 inhibits the kinase activity of the cyclin D-CDK4 complex.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Platform
Microplate
Properties
Storage instructions
Store at +4°C. Please refer to protocols.Components 1 x 96 tests 10X Human p21 Capture Antibody 1 x 600µl 10X Human p21 Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human p21 Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS(ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMBDevelopment Solution 1 x 12ml Research areas
- Cell Biology
- Cell Cycle
- Cell Cycle Inhibitors
- Cip / Kip
- Epigenetics and Nuclear Signaling
- DNA / RNA
- DNA Damage & Repair
- DNA Damage Response
- p53
- Epigenetics and Nuclear Signaling
- Cell cycle
- Cell Cycle Inhibitors
- Cip/Kip
- Cancer
- Cell cycle
- Cell cycle inhibitors
- Cip/kip
- Cancer
- Oncoproteins/suppressors
- Tumor suppressors
- p53 pathway
- Kits/ Lysates/ Other
- Kits
- ELISA Kits
- ELISA Kits
- Cancer proteins ELISA kits
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-translationalmodifications
Phosphorylation 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
Alternative names
- CAP20
- CDK-interacting protein 1
- CDKI
see allDatabase links
- Entrez Gene:1026 Human
- Omim:116899 Human
- SwissProt:P38936 Human
- Unigene:370771 Human
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