Mouse p21 ELISA Kit (CDKN1A) (ab205576)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 2.7 pg/ml
- Range: 62.5 pg/ml - 4000 pg/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
Overview
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Product name
Mouse p21 ELISA Kit (CDKN1A)
See all p21 kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Cell extract 5 3.4% Inter-assay Sample n Mean SD CV% Cell extract 3 7.8% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
2.7 pg/ml -
Range
62.5 pg/ml - 4000 pg/ml -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Mouse -
Product overview
Mouse p21 ELISA Kit (CDKN1A) (ab205576) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of p21 (CDKN1A) protein in cell culture extracts and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse p21 (CDKN1A) with 2.7 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 stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
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. p21 may be the important intermediate by which p53/TP53 mediates its role as an inhibitor of cellular proliferation in response to DNA damage.
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
Pre-coated microplate (12 x 8 well strips)
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Mouse p21 Capture Antibody 1 x 600µl 10X Mouse 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 5BR 1 x 6ml Mouse 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 TMB Development Solution 1 x 12ml -
Research areas
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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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Alternative names
- CAP20
- CDK-interacting protein 1
- CDKI
see all -
Database links
- Entrez Gene: 12575 Mouse
- SwissProt: P39689 Mouse
- Unigene: 195663 Mouse
Images
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Other - Mouse p21 ELISA Kit (CDKN1A) (ab205576)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of p21 standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of p21 in mouse NIH/3T3 cell extract samples.The concentrations of p21 were measured in duplicates, interpolated from the p21 standard curves and corrected for sample dilution. Note that 1X Diluted NIH/3T3 cell extract samples were at 80 µg/mL. The interpolated, dilution factor-corrected values are plotted in pg of p21 per µg of total protein (mean +/- SD, n=2).
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Camptothecin treatment of NIH/3T3 cells induces expression of p21.NIH/3T3 cells were cultured in the absence or presence of 1 µM camptothecin for 18 hours. Raw 264.7 cells were cultured in the absence of camptothecin. The cell extracts were prepared. The concentrations of p21 were measured in the diluted cell extracts in duplicates, interpolated from the p21 standard curves and corrected for sample dilution. Note that 1X Diluted untreated NIH/3T3 cell extract samples were at 44 µg/mL. Note that 1X Diluted camptothecin treated NIH/3T3 cell extract samples were at 11 µg/mL. Note that 1X Diluted untreated Raw 264.7 cell extract samples were at 6 µg/mL. The interpolated, dilution factor-corrected values are plotted in pg of p21 per µg of total protein (mean +/- SD, n=2).
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Sandwich ELISA - Mouse p21 ELISA Kit (CDKN1A) (ab205576)To learn more about the advantages of recombinant antibodies see here.
