Anti- CDT2 / DTL antibody, rabbit serum
Product code70-115

Size

100µl

Storage

Store at 4℃ for short term. For long term storage store at -20℃.
Aliquot to avoid repeated freezing thawing.

Concentration

N/ABuffer

0.05 % sodium azide

PurityRabbit antiserum

ImmunogenE. coli expressed and purified 6×His-tagged C-terminal 150 amino acids ofhuman Cdt2

IsotypeRabbit IgG

Reactivity

Human and mouse, rat, hamster

Special notes

Specificity was validated by western blotting with siRNA (Fig.2)

Application

1. Western blotting (1/200-1/2,000 dilution).
2. Immunoprecipitation (assay dependent)
3. Immunofluorescence staining (1/100~1/1,000 dilution)
4. Flow Cytometry (assay dependent).
Other applications have not been tested.

BackgroundCDT2 (human, 730 aa, 79.5 kDa) is substrate-specific adapter of a DCX (DDB1CUL4-X-box) E3 ubiquitin-protein ligase complex required for cell cycle control,
DNA damage response and translesion DNA synthesis. The DCX(DTL) complex,
also named CRL4(CDT2) complex, mediates the polyubiquitination andsubsequent degradation of CDT1 and CDKN1A/p21(CIP1). CDT1 degradationin response to DNA damage is necessary to ensure proper cell cycle regulationof DNA replication. CDKN1A/p21(CIP1) degradation during S phase orfollowing 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 containing the 'K+4'
motif in the PIP box, recruit the DCX(DTL) complex, leading to theirdegradation. In undamaged proliferating cells, the DCX(DTL) complex also
promotes the 'Lys-164' monoubiquitination of PCNA, thereby being involved inPCNA-dependent translesion DNA synthesis. CDT2 is activated by checkpoint
kinase ATR following DNA damage.

Data Link

UniProtKBQ9NZJ0 Human

Entrez Gene: 51514 Human

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: 70-117 Anti- CDT2 / DTL antibody, rabbit serumFig.1. 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.

Fig.2. Inhibition of Cdt2 protein synthesis by Cdt2 siRNA introduced into HeLa cells. siCont is control
siRNA unrelated to Cdt2. siCdt2 is Cdt2 specific siRNA.
Cdt2 is phosphorylated after UV irradiation as shown by the band shift-up in irradiated sample (UV+).
References: This product was described in Ref.1 and used in the following publications.
1. Nishitani H. et al. (2008) CDK Inhibitor p21 Is Degraded by a Proliferating Cell Nuclear Antigencoupled Cul4-DDB1Cdt2 Pathway during S Phase and after UV Irradiation. J.Biol.Chem. 283:29045-29052. WB Link
2. Ishii T. et al. (2010) Proliferating cell nuclear antigen-dependent rapid recruitment of Cdt1 and2 / 3

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

CRL4Cdt2 at DNA-damaged sites after UV irradiation in HeLa cells. J Biol Chem. 285:41993-42000.
WB, IF, FACS

Link

3. Roukos V. et al. (2011) Dynamic recruitment of licensing factor Cdt1 to sites of DNA damage. J.
Cell Science 124: 422-434. IF Link
4. Sakaguchi H. et al. (2012) Checkpoint Kinase ATR Phosphorylates Cdt2, a Substrate Receptor ofCRL4 Ubiquitin Ligase, and Promotes the Degradation of Cdt1 following UV Irradiation. PLoSONE 7(9): e46480. WB Link
5. Shiomi Y. et al. (2012) Two Different Replication Factor C Proteins, Ctf18 and RFC1, Separately
Control PCNA-CRL4Cdt2-Mediated Cdt1 Proteolysis during S Phase and following UV Irradiation.
Mol. Cell. Biol. 32: 2279-2288. WB, IF Link

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