概述

  • 产品名称Anti-CDT1抗体
    参阅全部 CDT1 一抗
  • 描述
    兔多克隆抗体to CDT1
  • 经测试应用适用于: WB, ICC/IFmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 500 to the C-terminus of Human CDT1.

    (Peptide available as ab93742.)

  • 阳性对照
    • This antibody gave a positive signal in both human brain and liver tissue lysates.

性能

应用

Our Abpromise guarantee covers the use of ab83174 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Detects a band of approximately 73 kDa (predicted molecular weight: 60 kDa).
ICC/IF Use a concentration of 5 µg/ml.

靶标

  • 功能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.
  • 序列相似性Belongs to the Cdt1 family.
  • 发展阶段Present during G1 and early S phase of the cell cycle. Degraded during the late S, G2, and M phases.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

Anti-CDT1 antibody 图像

  • All lanes : Anti-CDT1 antibody (ab83174) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human brain tissue lysate - total protein (ab29466)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 60 kDa
    Observed band size : 73 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 1 minute
  • ICC/IF image of ab83174 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab83174, 5µg/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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

Anti-CDT1 antibody (ab83174)参考文献

This product has been referenced in:
  • Jimenez-Hernandez M  et al. Exploring the spectroscopic differences of Caki-2 cells progressing through the cell cycle while proliferating in vitro. Analyst N/A:N/A (2013). Read more (PubMed: 23640135) »
  • Wong VC  et al. S-phase sensing of DNA-protein crosslinks triggers TopBP1-independent ATR activation and p53-mediated cell death by formaldehyde. Cell Cycle 11:2526-37 (2012). Read more (PubMed: 22722496) »

See all 2 Publications for this product

Product Wall

Abreviews
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 25°C
Sample Human Cell (Caki-2 (Clear cell renal carcinoma))
Specification Caki-2 (Clear cell renal carcinoma)
Permeabilization Yes - 0.25% X-Triton in PBS
Fixative Formaldehyde
Username

Dr. Melody Jimenez

Verified customer

提交于 Feb 11 2013

I think its good that you are checking the cell viability and using HeLa may provide more information as to if it is the cell strain or the antibodies that may be contributing to the problems. I agree that checking both the primary and secondary antibo...

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Thank you for providing such a full explanation of what you are trying to achieve. I can now understand exactly why you have chosen the technique that you have.

I think the experiments you are planning to do next will be very useful in determ...

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Thank you for getting back to me with your new results. It is good that we can now say conclusively that there is non-specific binding of both the secondary antibody and of the isotype control antibody.

Following the protocol changes, I do no...

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Thank you for that last bit of information.


I just wanted to check that the isotype control wasn't being used in a higher concentration than the primary antibody, but everything is fine.


I look forward to hearing how you...

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Sorry for the delay in getting back to you and thank you for all that information. It makes it much easier to understand what is going on and what has been tried previously.

I think the next step you are suggestingare good. The blocking buffe...

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Thank you for contacting us and reporting the problems you have encountered in using the Anti-CDT1 antibody (ab83174) and rabbit IgG isotype control (ab37406).


As discussed over the phone, in order to provide the most suitable suggestio...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100
Username

Dr. Kirk McManus

Verified customer

提交于 Mar 15 2011

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