Cell Signaling Technology

Product Pathways - Screening Technologies

Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb #5243

Applications Reactivity Sensitivity Isotype
W IP E-P All Endogenous Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  E-P=ELISA (Peptide)
Reactivity Key: All=All species expected
Species cross-reactivity is determined by western blot.

Protocols

Specificity / Sensitivity

Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb detects endogenous levels of cellular proteins containing the Ser-phospho-Thr-Pro motif. This antibody has minor cross-reactivity with phospho-Thr-Pro motifs.

Source / Purification

Monoclonal antibody is produced by immunizing animals with an SpTP peptide library.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or nocodazole-treated (100 ng/ml, 16 hrs), using Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb.

IP

IP

Immunoprecipitation of extracts from HeLa cells, untreated or nocodazole-treated (100 ng/ml, 16 hrs), using Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb (lanes 1 and 2). 10% input is shown in lanes 3 and 4. Western blots were performed using Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb (upper), RIP (D94C12) XP® Rabbit mAb #3493 (upper middle), MEK1/2 Antibody #9122 (lower middle), and NPM Antibody #3542 (lower).

Background

Polo-like kinases (PLKs) are Ser/Thr protein kinases that play an essential role during the cell cycle. At least four PLKs exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4 (1). PLKs have a highly conserved amino-terminal kinase domain and a relatively divergent carboxy-terminal domain called the Polo-box domain (PBD). Of the four PLKs, PLK1 is the best characterized (2). PLK1 functions as a key regulator of mitotic events by phosphorylating substrate proteins on centrosomes, kinetochores, the mitotic spindle, and the midbody, and is crucial for proper progression through multiple stages of mitosis (2-4). The PBDs of PLK1 function as a phospho-Ser/Thr-binding module, recognizing the optimal recognition sequence motif Ser-(pSer/pThr)-(Pro/X) (5). Binding of phosphopeptides containing the Ser-(pSer/pThr)-(Pro/X) motif by the PBD in PLK1 relieves its inhibitory function on kinase activity (6). Ser-(pSer/pThr)-Pro peptides are phosphorylated by proline-directed kinases such as cyclin dependent kinases (CDKs). These findings imply that priming phosphorylations on substrates or docking proteins by other mitotic kinases such as CDKs may target PLK1 to its substrates and simultaneously activate its kinase activity. Phospho-PLK Binding Motif (ST*P) (D73F6) Rabbit mAb is a useful tool to study PLK1 binding proteins and PLK1 substrates.

  1. Nigg, E.A. (1998) Curr Opin Cell Biol 10, 776-83.
  2. Donaldson, M.M. et al. (2001) J Cell Sci 114, 2357-8.
  3. Barr, F.A. et al. (2004) Nat Rev Mol Cell Biol 5, 429-40.
  4. Kishi, K. et al. (2009) Mol Cell Biol 29, 3134-50.
  5. Elia, A.E. et al. (2003) Science 299, 1228-31.
  6. Elia, A.E. et al. (2003) Cell 115, 83-95.

Application References

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License/Use Restrictions: Use of CST Motif Antibodies within certain methods (e.g., U.S. Patent No.’s 7,198,896 & 7,300,753) may require a license from CST. For information regarding academic licensing terms please have your technology transfer office contact CST Legal Department at CST_ip@cellsignal.com. For information regarding commercial licensing terms please contact CST Pharma Services Department at ptmscan@cellsignal.com.

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

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