Overview

  • Product name

    Anti-p21 (phospho T145) antibody
    See all p21 primary antibodies
  • Description

    Rabbit polyclonal to p21 (phospho T145)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, ELISAmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    synthesized phosphopeptide derived from human p21Cip1 around the phosphorylation site of threonine 145 (R-Q-TP-S-M)

  • Positive control

    • Human breast carcinoma tissue and EGF treated HeLa cell extracts
  • General notes


    p21Cip1 (phospho-Thr145) antibody detects endogenous levels of p21Cip1 only when phosphorylated at threonine 145.

Properties

Applications

Our Abpromise guarantee covers the use of ab47300 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
ICC/IF Use a concentration of 1 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 32 kDa (predicted molecular weight: 18 kDa).
IHC-P Use at an assay dependent concentration.
ELISA 1/10000.

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 (phospho T145) antibody (ab47300)

    Lane 1 : EGF treated HeLa cells
    Lane 2 : HeLa cells

    Predicted band size: 18 kDa

  • ab47300 staining human breast carcinoma tissue by IHC-P (left hand panel). The right hand panel shows staining in the presence of phospho-peptide.
  • ICC/IF image of ab47300 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47300, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:

See all 5 Publications for this product

Customer reviews and Q&As

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Skin)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Antigen retrieval (Dako)
Permeabilization
No
Specification
Skin
Fixative
Formaldehyde

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Submitted Nov 06 2017

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