Anti CDT2/DTL - Cosmo Bio Co.,Ltd.

Antibodies

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Anti CDT2/DTL Antibody

Catalog No.: BAM-70-115-EX
Size: 100UL
Price: ¥35000
$467
antigen/source: CDT2
host: Rabbit
Purity: Serum
Application: Western Blot
Immuno Fluorescence
Immunoprecipitation
Storage: -20C
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Immunogen: Human
Reacts with: Human
Mouse
Purity: Affinity Purified
Purpose: Western blotting: 1/2,000 dilution
Immunoprecipitation (1/100-1/200 dilution)
Immunofluorescence staining (1/100-1/1,000)
FACS (assay dependent)
Other applications have not been tested.
Optimal dilutions/concentrations should be determined by the e

Other:

 Background 
CDT2 (human, 730 aa, 79.5 kDa) is substrate-specific adapter of a DCX (DDB1-CUL4-X-box) E3 ubiquitin-protein ligase complex required for cell cycle control, DNA damage response and translesion DNA synthesis. The DCX(DTL) complex, alsonamed CRL4(CDT2) complex, mediates the polyubiquitination and subsequent degradation of CDT1 and CDKN1A/p21(CIP1). CDT1 degradation in response to DNA damage is necessary to ensure proper cell cycle regulation of DNA replication. CDKN1A/p21(CIP1) degradation during S phase or following UV irradiation is essential to control replication licensing. Most substrates require their interaction with PCNA for their polyubiquitination: substrates interact with PCNA via their PIP-box, and those containingthe 'K+4' motif in the PIP box, recruit the DCX(DTL) complex, leading to their degradation. In undamaged proliferating cells, the DCX(DTL) complex also promotes the 'Lys-164' monoubiquitination of PCNA, thereby being involved in PCNA-dependent translesion DNA synthesis. CDT2 is activated by checkpoint kinase ATR following DNA damage.
 Cross reactivity 
Human and mouse Cdt2 proteins. Not tested in other species



 IMAGE1



Identification of Cdt2 protein in whole cell extract of HeLa cells by western blotting. With cell extract of 20 µg protein, endogenous level of Cdt2 protein was detected with the antibody at 1/2,000 dilution. Secondary antibody was HRP-conjugated goat anti-rabbit IgG antibody at 1/20,000.

 IMAGE2



Inhibition of Cdt2 protein synthesis by Cdt2 siRNA introduced into HeLa cells. siCont is control siRNA unrelated to Cdt2. siCdt2 is Cdt2 siRNA. Cdt2 is phosphorylated after UV irradiation as shown by the band shift-up in irradiated sample (UV +).

 IMAGE3



Immunofluorescence staining of Cdt2 protein in growing Hela cells with the antibody. Asynchroneously growing HeLa cells were stained with Hoechst 33258 for DNA and with the antibody for Cdt2 protein. The cells were fixed in 4% paraformaldehyde for 10 min, permeabilized in 0.25% (v/v) Triton X-100 in phosphate-buffered saline (PBS), and reacted with the primary antibody. Alexa-592 conjgated goat anti-rabbit IgG was used as secondary antibody.Cdt2 protein is localized in nuclei at all cell stages.

Reference:

•  Nishitani H. et al., J.Biol.Chem. 283 3:29045-29052 (2008)
•  Ishii T. et al., J Biol Chem. 285 41993-42000 (2010)
•  Roukos V. et al., J. Cell Science 124 422-434 (2011)
•  Sakaguchi H. et al., PLoS ONE.7(9)
 
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