Cell Signaling Technology

Product Pathways - Protein Stability

Skp2 (L70) Antibody #4313

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC H M R Mk Endogenous 48 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
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

Skp2 (L70) Antibody detects endogenous levels of Skp2 protein (α, β, and γ isoforms). The antibody does not cross-react with other Skp proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu70 of human Skp2 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 Skp2 (L70) Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Skp2 (L70) Antibody (green). Actin filaments have been labeled with DY-554 phalloidin (red).

Background

Members of the F-box family of proteins are characterized by the approximate 40 amino acid F-box motif named after cyclin F (1,2). F-box proteins constitute one of the four subunits of the Skp1-Cullin-F-box (SCF) ubiquitin ligase complex. The substrate specificity of SCF complexes is determined by the interchangeable F-box proteins, which act as adaptors by associating with phosphorylated substrate proteins and recruiting them to the SCF core. F-box proteins contain two fundamental domains: the F-box motif mediates binding to Skp1 and a leucine rich repeat (LRR) domain mediates substrate interactions.

Skp2 (S phase kinase-associated protein 2) interacts with cyclin A/CDK2 and mediates proper G1 to S and G2 to M phase transitions by targeting the cyclin-dependent kinase (CDK) inhibitors p27, p21, p130 and the FoxO1 transcription factor for ubiquitylation and subsequent proteolysis (3-6). Skp2 protein expression is low in G0 and early G1 phase, increases during late G1 phase and peaks during S and G2 phases. Inactivation of Skp2 results in S/G2-phase arrested cells with endoduplication and multiple centrosomes (4). Overexpression of Skp2 results in increased CDK activity and contributes to the deregulated proliferation and genetic instabilities typical of cancer cells (7). Increased Skp2/decreased p27 levels are associated with many aggressive lymphomas and human carcinomas such as colon, breast, prostate and lung cancers (7).

  1. Pagano, M. (2004) Mol Cell 14, 414-416.
  2. Reed, S.I. (2003) Nature Rev Mol Cell Biol 4, 855-864.
  3. Zhang, H. et al. (1995) Cell 82, 915-25.
  4. Nakayama, K. et al. (2004) Dev Cell 6, 661-72.
  5. Bornstein, G. et al. (2003) J Biol Chem 278, 25752-7.
  6. Tedesco, D. et al. (2002) Genes Dev 16, 2946-57.
  7. Bloom, J. and Pagano, M. (2003) Semin Cancer Biol 13, 41-7.

Application References

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Companion Products

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

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