| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| Geminin (E5Q9S) XP® Rabbit mAb | 52508 | 20 µl | 25 kDa | Rabbit IgG |
| CDT1 (D10F11) Rabbit mAb | 8064 | 20 µl | 65 kDa | Rabbit IgG |
| Thymidine Kinase 1 (E2H7Z) Rabbit mAb | 28755 | 20 µl | 26 kDa | Rabbit IgG |
| Phospho-Histone H3 (Ser10) (D7N8E) XP® Rabbit mAb | 53348 | 20 µl | 17 kDa | Rabbit IgG |
| Cyclin A2 (E1D9T) Rabbit mAb | 91500 | 20 µl | 55 kDa | Rabbit IgG |
| Cyclin B1 (D5C10) XP® Rabbit mAb | 12231 | 20 µl | 55 kDa | Rabbit IgG |
| Cyclin E1 (D7T3U) Rabbit mAb | 20808 | 20 µl | 48 kDa | Rabbit IgG |
| Phospho-cdc2 (Tyr15) (10A11) Rabbit mAb | 4539 | 20 µl | 34 kDa | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat |
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Description
The Cell Cycle Phase Determination Antibody Sampler Kit provides an economical means of detecting total proteins or post-translational modifications present in cells at various phases of the cell cycle. Geminin is degraded in G1 phase, while CDT1 is degraded in S, G2, and M phases. Thymidine Kinase 1 accumulates in G1 phase, peaks in S phase, and is degraded before cell division. Phospho-Histone H3 (Ser10) is present only in M phase, while Phospho-cdc2 (Tyr15) is absent in M phase. Cyclins A2, B1, and E1 peak at G2 phase, late G2/M phase, and late G1/early S phase, respectively. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.
Storage
Background
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).
- Atherton-Fessler, S. et al. (1994) Mol Biol Cell 5, 989-1001.
- Gong, D. and Ferrell, J.E. (2010) Mol Biol Cell 21, 3149-61.
- Norbury, C. et al. (1991) EMBO J 10, 3321-9.
- Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
- Pagano, M. et al. (1992) EMBO J 11, 961-71.
- Furuno, N. et al. (1999) J Cell Biol 147, 295-306.
- Munch-Petersen, B. (2010) Nucleosides Nucleotides Nucleic Acids 29, 363-9.
- Caillat, C. and Perrakis, A. (2012) Subcell Biochem 62, 71-87.
Background References
Trademarks and Patents
Limited Uses
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