Cell Signaling Technology

Product Pathways - Protein Stability

Phospho-Skp2 (Ser64) Antibody #14865

E3 Ubiquitin Ligases   F-Box proteins   FBXL1   p-Skp2   Phospho-Skp2   SCF Ubiquitin Ligases   Skp2  

No. Size Price
14865S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
14865 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 48 Rabbit
IP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation,

Specificity / Sensitivity

Phospho-Skp2 (Ser64) Antibody recognizes endogenous levels of Skp2 protein only when phosphorylated at Ser64. This antibody also cross-reacts with an unidentified 160 kDa phosphoprotein in some cells.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser64 of human Skp2 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing DDK-tagged full-length, human Skp2 wild-type protein (hSkp2-WT-DDK; +) or DDK-tagged full-length, human Skp2 S64A protein (hSkp2-S64A-DDK; +), using Phospho-Skp2 (Ser64) Antibody (upper) and DYKDDDDK Tag Antibody #2368 (lower).



Immunoprecipitation of phosphorylated Skp2 from 293T cell extracts using Normal Rabbit IgG #2729 (lane 2) or Phospho-Skp2 (Ser64) Antibody (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Phospho-Skp2 (Ser64) Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells expressing either non-targeting shRNA (shNT) or Skp2 shRNA (shSkp2) using Phospho-Skp2 (Ser64) Antibody (upper), Skp2 (D3G5) XP® Rabbit mAb #2652 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T and HeLa cells using Phospho-Skp2 (Ser64) Antibody. The phospho-specificity of the antibody was verified by pre-incubating the antibody with no peptide (left), Skp2 non-phosphopeptide (center), and Skp2 (Ser64) phosphopeptide (right).


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.

S phase kinase-associated protein 2 (Skp2) interacts with cyclin A/CDK2 and mediates G1 to S and G2 to M phase transitions by targeting the cyclin-dependent kinase (CDK) inhibitors p27, p21, and p130 for ubiquitination and subsequent proteolysis (3-6). Overexpression of Skp2 results in deregulated proliferation and genetic instabilities typical of cancer cells (7). Research studies have shown that increased Skp2/decreased p27 levels are associated with many aggressive lymphomas and human carcinomas such as colon, breast, prostate and lung cancers (7). Several recent research studies have demonstrated that Skp2 is subject to phosphorylation-dependent regulation by a network of pro-proliferative Ser/Thr kinases. It appears as though phosphorylation of Skp2 at Ser64 by CDK2 (8), Ser72 by Akt1 (9), and Thr417 by PIM1 (10) promotes stabilization of Skp2, possibly constituting an additional mechanism for Skp2 oncogenicity.

  1. Pagano, M. (2004) Mol Cell 14, 414-6.
  2. Reed, S.I. (2003) Nat Rev Mol Cell Biol 4, 855-64.
  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.
  8. Rodier, G. et al. (2008) EMBO J 27, 679-91.
  9. Gao, D. et al. (2009) Nat Cell Biol 11, 397-408.
  10. Cen, B. et al. (2010) J Biol Chem 285, 29128-37.

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