Overview

  • Product name
  • Description
    Rabbit polyclonal to CDT1
  • Tested applications
    Suitable for: IP, IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Chimpanzee
  • Immunogen

    A region between residue 500 and the C-terminus (residue 546) of human CDT1 (NP_112190.1).

  • Positive control
    • HeLa and 293T whole cell lysates.

Properties

Applications

Our Abpromise guarantee covers the use of ab70829 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
IP Use at 2-5 µg/mg of lysate.
IHC-P 1/1000 - 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
WB 1/1000 - 1/2500. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).

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

  • All lanes : Anti-CDT1 antibody (ab70829) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate at 50 µg
    Lane 2 : HeLa whole cell lysate at 15 µg
    Lane 3 : HeLa whole cell lysate at 5 µg
    Lane 4 : 293T whole cell lysate at 50 µg

    Developed using the ECL technique

    Predicted band size : 63 kDa
    Observed band size : 63 kDa


    Exposure time : 3 minutes

    Secondary antibody - anti-rabbit HRP (ab6721)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human islet cell carcinoma tissue labelling CDT1 with ab70829 at 1/1000 (1µg/ml). Detection: DAB.
  • Detection of CDT1 by Western Blot of Immunprecipitate.
    ab70829 at 1µg/ml staining CDT1 in HeLa whole cell lysate immunoprecipitated using ab70829 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded/lane).

    Secondary antibody anti-rabbit HRP (ab6721)
    Detection: Chemiluminescence with exposure time of 3 minutes.

References

This product has been referenced in:
  • Hernández-Pérez S  et al. USP37 deubiquitinates Cdt1 and contributes to regulate DNA replication. Mol Oncol N/A:N/A (2016). WB ; Human . Read more (PubMed: 27296872) »
  • Grandy RA  et al. Genome-Wide Studies Reveal that H3K4me3 Modification in Bivalent Genes Is Dynamically Regulated during the Pluripotent Cell Cycle and Stabilized upon Differentiation. Mol Cell Biol 36:615-27 (2015). WB ; Human . Read more (PubMed: 26644406) »

See all 7 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa S3 cervical cancer)
Loading amount
50 µg
Specification
HeLa S3 cervical cancer
Treatment
double thymidine block and release
Gel Running Conditions
Reduced Denaturing (8% Tris-glycine SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted May 05 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SAOS-2 OSTEOSARCOMA)
Specification
SAOS-2 OSTEOSARCOMA
Fixative
Paraformaldehyde
Permeabilization
Yes - PBS TRITON .25%
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 03 2012

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (embryonic stem cells)
Specification
embryonic stem cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton 0.5%
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
Username

Mr. Shai Melcer

Verified customer

Submitted Nov 18 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Balb MK/ mouse keratinocyte / and 3T3 / and CHO)
Loading amount
500000 cells
Specification
Balb MK/ mouse keratinocyte / and 3T3 / and CHO
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Mr. Thinh Cao

Verified customer

Submitted May 07 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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