Technical Data
C2562-95
Cdc45
Description:
The initiation of DNA replication in mammalian cells is a highly coordinated process that ensures duplication of the genome only once per cell division cycle. Origins of replication are dispersed throughout the genome and their activities are regulated via the sequential binding of pre-replication and replication factors. The origin recognition complex (ORC) is thought to bind to chromatin throughout the cell cycle (1,2). The pre-replication complex (Pre-RC) forms in late mitosis/early G1 phase with the binding of CDT1 and cdc6 to the origin, which allows binding of the heterohexameric MCM2-7 complex. The MCM complex is thought to be the replicative helicase and formation of the pre-RC is referred to as chromatin licensing. Subsequent initiation of DNA replication requires the activation of the S-phase promoting kinases cdk2 and cdc7. Cdc7 phosphorylates MCM proteins bound to chromatin and, in conjunction with CDT1, recruits the replication factor cdc45 (3-5). Cdc45 is required for formation of pre-initiation com- plexes at the G1/S transition and for activation of replication origins. The level of cdc45 protein expression is associated with the proliferative status of the cell or tissue. Terminally differentiated and senescent cells lack cdc45 and highly proliferative cell lines express high levels of cdc45 (6).

Applications:
Suitable for use in Western Blot. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000. Incubate membrane with diluted antibody in 5% w/v BSA, 1X TBS, 0.1%
Tween-20 at 4C with gentle shaking, overnight.
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ul-20CBlue IceHumanRabbit
Concentration:
Not determined
Immunogen:
Synthetic peptide corresponding to the carboxy terminus of human cdc45 protein (KLH-coupled).
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied in 10mM sodium HEPES pH 7.5, 150mM NaCl, 100ug/ml BSA and 50% glycerol.
Specificity:
Recognizes human Cdc45. Species crossreactivity: mouse, rat, monkey.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Okuno, Y. et al. (2001) EMBO J 20, 4263-77, 2. McNairn, A.J. et al. (2005) Exp Cell Res 308, 345-56, 3. Bell, S.P. and Dutta, A. (2002) Annu Rev Biochem 71, 333-74, 4. Tsuji, T. et al. (2006) Mol Biol Cell 17, 4459-72, 5. Ballabeni, A. et al. (2009) J Biol Chem 284, 3028-36, 6. Pollok, S. et al. (2007) FEBS J 274, 3669-84.