Name: Anti-p21 antibody [EPR18021]
Brand: RabMAb
SKU: ab188224
Rating: 5 (5 reviews)

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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
Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
Reacts with: Mouse

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Primary

Secondary

Protein

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Primary - Rabbit Anti-p21 antibody [EPR18021] (ab188224)
WB, IHC-P, Flow Cyt, IP, ICC/IF

Secondary - Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
IHC-P, WB, ELISA, IP

Protein - Recombinant Human p21 protein (ab56278)
Functional Studies, SDS-PAGE, WB

View more associated products

Product name

Anti-p21 antibody [EPR18021]
See all p21 primary antibodies
Description

Rabbit monoclonal [EPR18021] to p21
Host species

Rabbit
Specificity

Expression 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 applications

Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
Species reactivity

Reacts with: Mouse
Immunogen

Recombinant full length protein within Mouse p21 aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P39689
Positive control
- WB: RAW 264.7 whole cell lysate; NIH/3T3, untreated and treated with 1µM staurosporine for 2hrs, whole cell lysates. IHC-P: Mouse testis and lung tissues. ICC/IF: RAW 264.7 and NIH/3T3 cells. Flow Cyt: RAW 264.7 cells. IP: NIN/3T3 whole cell lysate.
General notes
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR18021
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human p21 protein (ab56278)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Our Abpromise guarantee covers the use of ab188224 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt		1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP		1/30.
ICC/IF		1/500.

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
modifications

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.
Target information above from: UniProt accession P38936 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 12575 Mouse
- SwissProt: P39689 Mouse
- Unigene: 195663 Mouse
Alternative names
- CAP20 antibody
- CDK-interacting protein 1 antibody
- CDKI antibody
- CDKN1 antibody
- Cdkn1a antibody
- CDN1A_HUMAN antibody
- CIP1 antibody
- Cyclin Dependent Kinase Inhibitor 1A antibody
- Cyclin-dependent kinase inhibitor 1 antibody
- Cyclin-dependent kinase inhibitor 1A (P21) antibody
- Cyclin-dependent kinase inhibitor 1A (p21, Cip1) antibody
- DNA Synthesis Inhibitor antibody
- MDA-6 antibody
- MDA6 antibody
- Melanoma differentiation-associated protein 6 antibody
- Melanoma differentiation-associated protein antibody
- p21 antibody
- P21 protein antibody
- p21CIP1 antibody
- p21Cip1/Waf1 antibody
- p21WAF antibody
- PIC1 antibody
- SDI1 antibody
- SLC12A9 antibody
- WAF1 antibody
- Wild type p53 activated fragment 1 (WAF1) antibody
- Wild type p53 activated fragment 1 antibody
- Wildtype p53-activated fragment 1 antibody
see all

Western blot - Anti-p21 antibody [EPR18021] (ab188224)

All lanes : Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution

Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 1µM staurosporine for 2hrs, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 18 kDa
Observed band size: 18 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression level of p21 protein can be induced using staurosporine (protein kinase C inhibitor) (PMID:7677742).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nuclear staining on mouse testis is observed (PMID: 9170103).

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on RAW264.7 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-p21 antibody [EPR18021] (ab188224)

Flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling p21 with ab188224 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-p21 antibody [EPR18021] (ab188224)

Anti-p21 antibody [EPR18021] (ab188224) at 1/1000 dilution + RAW264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 18 kDa
Observed band size: 18 kDa

Exposure time: 8 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling p21 with ab188224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Sporadic nuclear staining on mouse lung is observed (PMID: 25333671).

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling p21 with ab188224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on NIH/3T3 cells.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Immunoprecipitation - Anti-p21 antibody [EPR18021] (ab188224)

p21 was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab188224 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab188224 at 1/500 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: NIH/3T3 whole cell lysate 10 µg (Input).

Lane 2: ab188224 IP in NIH/3T3 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab188224 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Publishing research using ab188224? Please let us know so that we can cite the reference in this datasheet.

ab188224 has been referenced in 37 publications.

Aschacher T et al. Long interspersed element-1 ribonucleoprotein particles protect telomeric ends in alternative lengthening of telomeres dependent cells. Neoplasia 22:61-75 (2020). PubMed: 31846834
Cai Y et al. Elimination of senescent cells by ß-galactosidase-targeted prodrug attenuates inflammation and restores physical function in aged mice. Cell Res N/A:N/A (2020). PubMed: 32341413
Chen X et al. Circular RNA circFBXW4 suppresses hepatic fibrosis via targeting the miR-18b-3p/FBXW7 axis. Theranostics 10:4851-4870 (2020). PubMed: 32308754
Sun L et al. Nicotine triggers islet ß cell senescence to facilitate the progression of type 2 diabetes. Toxicology 441:152502 (2020). PubMed: 32473187
Liu N et al. 3-Acetyldeoxynivalenol induces lysosomal membrane permeabilization-mediated apoptosis and inhibits autophagic flux in macrophages. Environ Pollut 265:114697 (2020). PubMed: 32454357

View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (Bone marrow)

Permeabilization

Yes - 0.05% TX-100

Specification

Bone marrow

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

Submitted Jan 01 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Mouse Tissue sections (kidney)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Permeabilization

Specification

kidney

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Fixative

10% formalin

Abcam user community

Verified customer

Submitted Jan 01 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Rat Tissue sections (Liver)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Permeabilization

Specification

Liver

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Fixative

10% formalin

Abcam user community

Verified customer

Submitted Jan 01 2019

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Permeabilization

Yes - 0.5% Triton X-100

Specification

HeLa

Fixative

Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Aug 10 2018

Application

Western blot

Sample

Mouse Cell lysate - whole cell (ameloblast)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

10 µg

Specification

ameloblast

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 23 2017

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Key features and details

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

Target

Function

Tissue specificity

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-p21 antibody [EPR18021]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-p21 antibody [EPR18021]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-p21 antibody [EPR18021]

Immunocytochemistry/ Immunofluorescence abreview for Anti-p21 antibody [EPR18021]

Western blot abreview for Anti-p21 antibody [EPR18021]

Post-translational
modifications