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

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-p21 antibody [EPR18021] (ab188224)
      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.

    • Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)
      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.

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

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

    • Immunocytochemistry/ Immunofluorescence - Anti-p21 antibody [EPR18021] (ab188224)
      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.

    • Flow Cytometry - Anti-p21 antibody [EPR18021] (ab188224)
      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.

    • Immunoprecipitation - Anti-p21 antibody [EPR18021] (ab188224)
      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 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) »
    • Xu T  et al. Oridonin enhancesin vitroanticancer effects of lentinan in SMMC-7721 human hepatoma cells through apoptotic genes. Exp Ther Med 14:5129-5134 (2017). Read more (PubMed: 29201227) »

    See all 3 Publications for this product

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

    Customer reviews and Q&As

    Submit a review

    Filter by Application

    Filter by Species

    Filter by Ratings

    2 Abreviews

    10
    Votes: Highest First

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

     Good
    Abreviews
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton X-100
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More
    abreview image
    Username

    Dr. Kirk Mcmanus

    Verified customer

    Submitted Aug 10 2018

    Western blot abreview for Anti-p21 antibody [EPR18021]

     Excellent
    Abreviews
    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
    Read More
    abreview image
    Username

    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"

    Sign up