Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Geminin Antibody #5165

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk Endogenous 25 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Geminin Antibody detects endogenous levels of total geminin protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly80 of human geminin protein. Antibodies are purified using protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell types using Geminin Antibody.

IP

IP

Immunoprecipitation of geminin from 293T cells using Geminin Antibody. Western blot was performed using the same antibody. Lane 1 contains 5% input.

Background

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 be bound to chromatin throughout the cell cycle (1,2). The pre-replication complex (pre-RC) forms in late mitosis/early G1 phase beginning with the binding of CDT1 and cdc6 to the origin, which allows binding of the heterohexameric MCM2-7 complex. Once this complex is formed, the origin is “licensed” for initiation of DNA replication. In order to ensure that replication occurs only once per cell cycle, geminin binds to and inhibits CDT1 during the S, G2 and M phases. This prevents the recruitment of the MCM complex to the origins of replication, which blocks the premature reformation of the Pre-RC. At the metaphase/anaphase transition, geminin is degraded by the anaphase-promoting complex (APC) allowing for the formation of new pre-RC (3,4).

  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. Montanari, M. et al. (2006) Cell Death Differ 13, 1052-6.
  4. McGarry, T.J. and Kirschner, M.W. (1998) Cell 93, 1043-53.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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