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Product Usage Information

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

APC3 Antibody recognizes endogenous levels of total APC3 protein. This antibody does not cross-react with either APC8/CDC23 or APC6/CDC16.

Species predicted to react based on 100% sequence homology: Hamster, Xenopus, Bovine, Dog, Pig, Guinea Pig, Horse

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human APC3 protein. Antibodies are purified by protein A and peptide affinity chromatography.

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 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 interact with UBE2S and UBE2C E2 enzymes, in part, via the RING-finger domain-containing subunit, APC11 (3-5). APC/C activity is also strictly dependent upon its association with multiple cofactors. For example, the related proteins, Cdc20 and Cdh1/FZR1, 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).

Anaphase-promoting complex subunit 3 (APC3), APC8, and APC6 are components of the tetratricopeptide (TPR) APC/C subcomplex (9). The presence of APC3 is required for binding of Cdh1/FZR1 to the APC/C. This suggests that APC/C is activated by an association between Cdh1/FZR1 with APC3 that enables APC/C to recognize the D-box of substrates (6,10). APC3 localizes to the centrosome and the mitotic spindle, suggesting that APC3 plays a critical role in the transition from metaphase to anaphase (11). Phosphorylation of APC3 at multiple sites during mitosis likely leads to structural changes within the APC/C by altering subunit interactions or changing affinity for molecules that transiently associate with the APC/C, such as Cdh1/FZR1 (12,13).

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.  Tugendreich, S. et al. (1993) Proc Natl Acad Sci U S A 90, 10031-5.

10.  Vodermaier, H.C. et al. (2003) Curr Biol 13, 1459-68.

11.  Tugendreich, S. et al. (1995) Cell 81, 261-8.

12.  Topper, L.M. et al. Cell Cycle 1, 282-92.

13.  Kraft, C. et al. (2003) EMBO J 22, 6598-609.

Entrez-Gene Id 996
Swiss-Prot Acc. P30260

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APC3 Antibody