Overview

  • Product name

    Anti-CDT1/DUP antibody [EPR17891]
    See all CDT1/DUP primary antibodies
  • Description

    Rabbit monoclonal [EPR17891] to CDT1/DUP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human CDT1/DUP aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H211

  • Positive control

    • WB: HeLa and HEK-293 cell lysates; Human fetal lung lysate. IHC-P: Human colonic carcinoma and cervix carcinoma tissues. ICC/IF: HeLa and HT-29 cells. IP: HEK-293 whole cell lysate.
  • General notes

     

     

     This product was previously labelled as CDT1

     

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab202067 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 60 kDa (predicted molecular weight: 60 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/100. PubMed: 28077461

Target

  • Function

    Cooperates with CDC6 to promote the loading of the mini-chromosome maintenance complex onto chromatin to form the pre-replication complex necessary to initiate DNA replication. Binds DNA in a sequence-, strand-, and conformation-independent manner. Potential oncogene.
  • Sequence similarities

    Belongs to the Cdt1 family.
  • Developmental stage

    Present during G1 and early S phase of the cell cycle. Degraded during the late S, G2, and M phases.
  • 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.
  • Post-translational
    modifications

    Ubiquitinated by the DCX(DTL) complex, also named CRL4(CDT2) complex, in response to DNA damage, leading to its degradation. Ubiquitination by the DCX(DTL) complex is necessary to ensure proper cell cycle regulation 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. The interaction with GMNN protects it against ubiquitination.
    Phosphorylated by cyclin A-dependent kinases which results in the binding of CDT1 to the F-box protein SKP2 and subsequent degradation. Binding to GMNN is not affected by phosphorylation.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • CDT 1 antibody
    • cdt1 antibody
    • CDT1_HUMAN antibody
    • Chromatin licensing and DNA replication factor 1 antibody
    • DNA replication factor antibody
    • DNA replication factor Cdt1 antibody
    • Double parked antibody
    • Double parked Drosophila homolog of antibody
    • Double parked homolog antibody
    • DUP antibody
    • Retroviral integration site 1 antibody
    • Retroviral integration site 2 antibody
    • Retroviral integration site1 antibody
    • Retroviral integration site2 antibody
    • RIS 2 antibody
    • RIS2 antibody
    see all

Images

  • Anti-CDT1/DUP antibody [EPR17891] (ab202067) at 1/10000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 60 kDa
    Observed band size: 60 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling CDT1/DUP with ab202067 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202067 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue labeling CDT1/DUP with ab202067 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. 

    Nuclear staining on human cervix carcinoma tissue is observed.

    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.

  • CDT1/DUP was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab202067 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab202067 at 1/1000 dilution.

    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG was used as secondary antibody at 1/1500 dilution.

    Lane 1: HEK-293 whole cell lysate 10 µg (Input).

    Lane 2: ab202067 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202067 in HEK-293 whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

  • Anti-CDT1/DUP antibody [EPR17891] (ab202067) at 1/2000 dilution + HEK-293 (Human epithelial cells from embryonic kidney) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 60 kDa
    Observed band size: 60 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human colonic carcinoma tissue labeling CDT1/DUP with ab202067 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on Human colonic carcinoma tissue is observed.

    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.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HT-29 (Human colorectal adenocarcinoma cells) cells labeling CDT1/DUP with ab202067 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

    Confocal image showing nuclear staining on T-29 cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab202067 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Anti-CDT1/DUP antibody [EPR17891] (ab202067) at 1/1000 dilution + Human fetal lung lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 60 kDa
    Observed band size: 60 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

  • Odagiri N  et al. Involvement of ERK1/2 activation in the gene expression of senescence-associated secretory factors in human hepatic stellate cells. Mol Cell Biochem N/A:N/A (2018). Read more (PubMed: 30426301) »
  • Zhang K  et al. Demethylzeylasteral inhibits glioma growth by regulating the miR-30e-5p/MYBL2 axis. Cell Death Dis 9:1035 (2018). Read more (PubMed: 30305611) »
See all 5 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (MCF10A epithelial cells)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
MCF10A epithelial cells
Blocking step
10% FBS, 1% BSA, 0.1% TX-100, 0.01% NaN3 in PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 27°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jan 23 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 Nov 03 2018

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