Overview

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.

Target

  • 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.
  • Information by UniProt
  • Database links
  • 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

Images

  • 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).

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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.

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

References

This product has been referenced in:
  • Jin Y  et al. Double deletion of tetraspanins CD9 and CD81 in mice leads to a syndrome resembling accelerated aging. Sci Rep 8:5145 (2018). WB . Read more (PubMed: 29572511) »
  • Jurisevic M  et al. The organic ester O,O'-diethyl-(S,S)-ethylenediamine-N,N'-di-2-(3-cyclohexyl)propanoate dihydrochloride attenuates murine breast cancer growth and metastasis. Oncotarget 9:28195-28212 (2018). Read more (PubMed: 29963272) »
See all 5 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

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

Abcam guarantees this product to work in the species/application used in this Abreview.
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 AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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