Cell Signaling Technology

Product Pathways - Protein Folding/Stability

Skp2 Antibody #4358

Applications Reactivity MW (kDa) Source
W IF-IC H Mk 48 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

This 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 rabbits with a synthetic peptide (KLH-coupled) corresponding to amino acids at the amino terminus of human Skp2 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, K562 and COS cells using Skp2 Antibody.

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells using Skp2 Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).

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 SCF (Skp1-Cullin-F-box) 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 (3,4,5,6). 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, 915-925.
  4. Nakayama, K. et al. (2004) Dev. Cell 6(5) , 661-672.
  5. Bornstein, G. et al. (2003) J. Biol. Chem. 278(28) , 25752-25757.
  6. Tedesco, D. et al. (2002) Genes Dev. 16(22) , 2946-2957.
  7. Bloom, J. and Pagano, M. (2003) Semin. Cancer Biol. 13(1) , 41-47.

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