Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

APC6 (D8D8) Rabbit mAb #9499

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Mk (B) (Dg) (Pg) Endogenous 72 Rabbit IgG

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

Protocols

Specificity / Sensitivity

APC6 (D8D8) Rabbit mAb recognizes endogenous levels of total APC6 protein. Based upon sequence alignment, this antibody is not predicted to cross-react with either APC8/CDC23 or APC3/CDC27.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human APC6 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human APC6 (hAPC6-Myc/DDK, +), using APC6 (D8D8) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using APC6 (D8D8) Rabbit mAb.

Background

Cell proliferation in all eukaryotic cells depends strictly upon the ubiquitin ligase (E3) activity of the anaphase promoting complex/cyclosome (APC/C), whose main function is to trigger the transition of the cell cycle from metaphase to anaphase. APC/C is a 1.5 MDa protein complex found in the nucleus of interphase cells. This complex diffuses throughout the cytoplasm and associates with parts of the spindle apparatus during mitosis. APC/C performs its various functions by promoting the assembly of polyubiquitin chains on substrate proteins, which targets these proteins for degradation by the 26S proteasome (1,2). In humans, twelve different APC/C subunits have been identified. Like all E3 enzymes, APC/C utilizes ubiquitin residues that have been activated by E1 enzymes and then transferred to E2 enzymes. Indeed APC/C has been shown to transiently interact with UBCH5 and UBCH10 E2 enzymes, in part, via the RING-finger domain-containing subunit, APC11 (3-5). In addition to E2 enzymes, APC/C activity is also strictly dependent upon one of several cofactors that associate with APC/C during specific phases of the cell cycle. The best studied of these are Cdc20 and Cdh1, which contain a C-terminal WD40 domain and participate in the recognition of APC/C substrates by interacting with specific recognition elements in these substrates (6), called D-Boxes (7) and KEN-boxes (8).

APC6/CDC16 is a component of the tetratricopeptide repeat (TPR) sub-complex of the APC/C, which also consists of APC8/CDC23 and APC3/CDC27. It is thought that this sub-complex plays an important role in coordinating the juxtaposition of the catalytic and substrate recognition modules relative to co-activator, regulatory proteins, and substrates (9). There is also evidence suggesting that phosphorylation of APC6 and the other TPR subunits during mitosis plays a functional role in regulating the association between TPR subunits and substrate recognition subunits such as Cdc20 (10).

  1. Qiao, X. et al. (2010) Cell Cycle 9, 3904-12.
  2. Harper, J.W. et al. (2002) Genes Dev 16, 2179-206.
  3. Carroll, C.W. and Morgan, D.O. (2002) Nat Cell Biol 4, 880-7.
  4. Gmachl, M. et al. (2000) Proc Natl Acad Sci U S A 97, 8973-8.
  5. Leverson, J.D. et al. (2000) Mol Biol Cell 11, 2315-25.
  6. Kraft, C. et al. (2005) Mol Cell 18, 543-53.
  7. Glotzer, M. et al. (1991) Nature 349, 132-8.
  8. Pfleger, C.M. and Kirschner, M.W. (2000) Genes Dev 14, 655-65.
  9. Schreiber, A. et al. (2011) Nature 470, 227-32.
  10. Kraft, C. et al. (2003) EMBO J 22, 6598-609.

Application References

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

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