Cell Signaling Technology

Product Pathways - Protein Stability

APC2 Antibody #12301

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk (Hm) (B) (Dg) (GP) Endogenous 100 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog  GP=Guinea 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

APC2 Antibody recognizes endogenous levels of total APC2 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Lys458 of human APC2 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using APC2 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human APC2 (hAPC2-Myc/DDK; +), using APC2 Antibody.

IP

IP

Immunoprecipitation of APC2 from 293T cell extracts, using Normal Rabbit IgG #2729 (lane 2) or APC2 Antibody (lane 3). Lane 1 is 10% input. Western blot analysis was performed using APC2 Antibody.


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 that has 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/FZR1, 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).

Anaphase-promoting complex subunit 2 (APC2) is a distant member of the cullin family (9,10) that interacts with a RING-H2 finger protein related to Rbx1/Hrt1/Roc1, called APC11, to form the catalytic subcomplex of the APC/C. The APC2/11 subcomplex recruits E2 enzymes such as UBE2C/UBCH10 and is required for the APC/C to catalyze substrate ubiquitination (11). Therefore, APC is a member of the expanding family of cullin-RING finger-based ubiquitin ligases. The physiologic importance of APC2 was underscored by the finding that disruption of murine Apc2 causes embryonic lethality (12).

  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. Zachariae, W. et al. (1998) Science 279, 1216-9.
  10. Yu, H. et al. (1998) Science 279, 1219-22.
  11. Tang, Z. et al. (2001) Mol Biol Cell 12, 3839-51.
  12. Wirth, K.G. et al. (2004) Genes Dev 18, 88-98.

Application References

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

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