|Geminin (E5Q9S) XP® Rabbit mAb 52508||20 µl||
||H Mk||25||Rabbit IgG|
|CDT1 (D10F11) Rabbit mAb 8064||20 µl||
||H Mk||65||Rabbit IgG|
|Thymidine Kinase 1 (E2H7Z) Rabbit mAb 28755||20 µl||
|Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit mAb 53348||20 µl||
||H M R Mk||17||Rabbit IgG|
|Cyclin A2 (E1D9T) Rabbit mAb 91500||20 µl||
|Cyclin B1 (D5C10) XP® Rabbit mAb 12231||20 µl||
||H R||55||Rabbit IgG|
|Cyclin E1 (D7T3U) Rabbit mAb 20808||20 µl||
||H M R||48||Rabbit IgG|
|Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb 4539||20 µl||
||H M R Mk||34||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding phosphorylated Ser10 of human histone H3 protein, phosphorylated Tyr15 of human cdc2 protein, Val421 of human cyclin A2 protein, Pro209 of human thymidine kinase 1 protein, Gly351 of human cyclin E1 protein, Pro70 of human geminin protein, residues near the amino terminus of human CDT1 protein, and residues near the amino terminus of human cyclin B1 protein.
The entry of eukaryotic cells into mitosis is regulated by cdc2/CDK1 kinase activation, a process controlled at several steps including cyclin B1 nuclear accumulation and binding, and phosphorylation of cdc2/CDK1 at Thr161 (1). At the end of mitosis, cyclin B1 is targeted for degradation by the anaphase-promoting complex (APC), allowing for cell cycle progression (2). A critical regulatory step in activating cdc2 during progression into mitosis is dephosphorylation of cdc2/CDK1 at Thr14 and Tyr15 (3).
Phosphorylation of Histone H3 at Ser10 is tightly correlated with chromosome condensation during both mitosis and meiosis (4).
Overcoming the G1/S checkpoint to commence DNA replication requires cyclin E, traversing the G2/M checkpoint to initiate mitosis requires cyclin B, and cyclin A is required for both S-phase and M-phase (5). Cyclin A availability is apparently the rate-limiting step for entry into mitosis, and cyclin A is required for completion of prophase (6).
Thymidine kinases play a critical role in generating the DNA synthetic precursor deoxythymidine triphosphate (dTTP). Cytoplasmic thymidine kinase 1 (TK1) expression and activity are regulated in a cell cycle-dependent manner, accumulating during G1-phase to peak levels in S-phase before being degraded prior to cell division (7).
The initiation of S phase begins with the formation of the pre-replication complex (pre-RC) in late mitosis/early G1 phase. CDT1 and cdc6 bind to the origin of DNA replication, which allows binding of the MCM2-7 complex. In order to ensure that replication occurs only once per cell cycle, geminin inhibits and destabilizes CDT1 during the S, G2 and M phases. At the metaphase/anaphase transition, geminin is degraded by the anaphase-promoting complex (APC) allowing for the formation of new pre-RC (8).
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