Anti-CDC6 antibody, rabbit polyclonal, ChIP-Grade
Product code70-133Size

100 µgStorage-20℃

Concentration

1.0 mg/ml

Buffer

PBS- with 50% glycerol. Filter-sterilized. Azide and carrier freePurity

The antiserum was first adsorbed with GST conjugated agarose column andthen the antibody in the pass-through fractions were purified with GST-CDC6conjugated column.

Immunogen

Recombinant GST-human CDC6 (1-326 amino acids) expressed in E. coli

IsotypeRabbit IgG

Reactivity

Human, mouse, rat

Special notes

Validation: Specificity of reaction has been validated with siRNA in WB (Ref.5& 7)

Application

1. Western blotting (1/1,000~3,000)
2. Immunoprecipitation （1/200）
3. Chromatin Immuno-Precipitation (assay dependent)
4. Immunofluorescence staining (1/200)

Background

CDC6 (Cell Division Control Protein 6 homolog) (human; 560 aa, 62.7 kDa) is
involved in the initiation of DNA replication. Also participates in checkpoint
controls that ensure DNA replication is completed before mitosis is initiated.

Data Link

uniprot/Q99741(CDC6_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: 70-133 Anti-CDC6 antibody, rabbit polyclonal, ChIP-Grade

Fig.1 Western blotting of endogenous CDC6 protein in HeLa cells
Lane 1; Size marker proteins (kDa)
Lane 2; HeLa cell whole lysate (10 μg)
Anti-CDC6 antibody was used at 1/1,000 dilution.

Fig 2. Immunoprecipitation of CDC6.
CDC6 protein was precipitated from whole cell lysate of HeLa cells with anti-CDC6 antibody beads
and probed by western blotting with anti-CDC6 antibody.
1; Control IP with non-immune IgG
2; Immunoprecipitates with anti-CDC6 antibody
3; Input (whole cell lysate)

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Fig.3 Immunofluorescence staining of CDC6 protein in HeLa cells
Confocal microscopic analyses of localization of CDC6 proteins in HeLa cells in G1 or early S phase.
Cells in mid to late G1phase or hydroxyurea-treated early S phase were fixed with formaldehyde
directly or after extraction with Triton X-100 and then immunostained with anti-CDC6 or control
non-immune antibody (Alexa;green). The samples were further treated with propidium iodide for
DNA staining (PI; red).

Fig.4 Immunofluorescence staining of CDC6 in MEF cells
MEFs (mouse embryonic fibroblasts) from embryonic day 14.5 mouse embryos were fixed with 10 %formalin at room temperature (RT) for 10 min, permeabilized with ice-cold methanol on ice for 10 min,and treated with 2 % BSA/PBS at RT for 1h. The cells were reacted with anti-cdc6 antibody (1:200)
at 4°C overnight and then with Alexa 555-conjugated goat anti-rabbit IgG antibody (1:1000) at RT for
1h. Chromosomal DNA was counterstained with 3.3 μM Hoechst 33342. Note that CDC6 abundantly
localizes in nuclei of the cells at G1 and early S phase.
Reference: This antibody was described in Ref.1 and used in the following publications.
1. Fujita M. et al. (1999) Cell cycle regulation of human CDC6 protein. Intracellular localization,interaction with the human mcm complex, and CDC2 kinase-mediated hyperphosphorylation. JBiol Chem. 274:25927-32. PMID:10464337, WB, IP, IF, (human)
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2. Fujita M. et al. (2002) Nuclear organization of DNA replication initiation proteins in mammaliancells. J Biol Chem. 277:10354-61. PMID: 11779870, WB, IP, IF, (human)
3. Tatsumi Y. et al. (2006) Deregulation of Cdt1 induces chromosomal damage without rereplicationand leads to chromosomal instability. J Cell Sci. 119:3128-40. PMID: 16835273. WB, (human)
4. Sugimoto N. et al. (2009) Redundant and differential regulation of multiple licensing factors
ensures prevention of re-replication in normal human cells. J Cell Sci. 15:1184-91.PMID:19339550,
WB, (human)
5. Yoshida K. et al. (2010) CDC6 interaction with ATR regulates activation of a replication checkpoint
in higher eukaryotic cells. J Cell Sci. 123:225-35. PMID:20048340. WB, IP, (human)6. Sugimoto N. et al. (2011) Chromatin remodeler sucrose nonfermenting 2 homolog (SNF2H) is
recruited onto DNA replication origins through interaction with Cdc10 protein-dependenttranscript 1 (Cdt1) and promotes pre-replication complex formation. J Biol Chem. 286:39200-10.
PMID:21937426. WB, (human)
7. Sugimoto N et al. (2015) Cdt1-binding protein GRWD1 is a novel histone-binding protein thatfacilitates MCM loading through its influence on chromatin architecture. Nucleic Acids Res. 2015
Jul 13;43(12):5898-911. PMID:25990725. WB, ChIP (human)

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