Cell Signaling Technology

Product Pathways - Cell Cycle / Checkpoint

Polo-like Kinase Antibody Sampler Kit #9785

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
PLK1 (208G4) Rabbit mAb #4513 40 µl W IP IHC-P H R Mk 62 Rabbit IgG
PLK3 (D14F12) Rabbit mAb #4896 40 µl W IP H M (R) (Mk) 80 Rabbit IgG
PLK4 Antibody #3258 40 µl W IF-IC H M R Mk 95 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

PLK1 (208G4) Rabbit mAb detects endogenous levels of of total PLK1 protein. PLK3 (D14F12) Rabbit mAb detects endogenous levels of total PLK3 protein. PLK4 Antibody detects endogenous levels of total PLK4 protein.

Western Blotting

Western Blotting

Western blot analysis of recombinant GST-tagged PLK4 Kinase #7580 and extracts from MOLT4, RL and Ramos cell lines using PLK4 Antibody #3258.

Western Blotting

Western Blotting

Western blot analysis of GSK-PLK1 fusion protein and extracts from HeLa, COS and PC12 cells, using PLK1 (208G4) Rabbit mAb #4513.

Western Blotting

Western Blotting

Western blot analysis of extracts from HEL, A549 and A20 cells using PLK3 (D14F12) Rabbit mAb #4896.


Description

The Polo-like Kinase Antibody Sampler Kit provides an economical means to investigate three distinct polo-like kinases within cells. The kit contains en ough primary and secondary antibodies to perform four western blot experiments per primary antibody.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human PLK1 or with a synthetic peptide corresponding to residues surrounding Cys625 of human PLK3 protein. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Cys458 of human PLK4. Antibodies are purified by peptide affinity chromatography.

Background

At least four distinct polo-like kinases exist in mammalian cells: PLK1, PLK2, PLK3, and PLK4/SAK (1). PLK1 apparently plays many roles during mitosis, particularly in regulating mitotic entry and exit. The mitosis promoting factor (MPF), cdc2/cyclin B1, is activated by dephosphorylation of cdc2 (Thr14/Tyr15) by cdc25C. PLK1 phosphorylates cdc25C at Ser198 and cyclin B1 at Ser133 causing translocation of these proteins from the cytoplasm to the nucleus (2-5). PLK1 phosphorylation of Myt1 at Ser426 and Thr495 has been proposed to inactivate Myt1, one of the kinases known to phosphorylate cdc2 at Thr14/Tyr15 (6). Polo-like kinases also phosphorylate the cohesin subunit SCC1, causing cohesin displacement from chromosome arms that allow for proper cohesin localization to centromeres (7). Mitotic exit requires activation of the anaphase promoting complex (APC) (8), a ubiquitin ligase responsible for removal of cohesin at centromeres, and degradation of securin, cyclin A, cyclin B1, Aurora A, and cdc20 (9). PLK1 phosphorylation of the APC subunits Apc1, cdc16, and cdc27 has been demonstrated in vitro and has been proposed as a mechanism by which mitotic exit is regulated (10,11).Substitution of Thr210 with Asp has been reported to elevate PLK1 kinase activity and delay/arrest cells in mitosis, while a Ser137Asp substitution leads to S-phase arrest (12). In addition, while DNA damage has been found to inhibit PLK1 kinase activity, the Thr210Asp mutant is resistant to this inhibition (13). PLK1 has been reported to be phosphorylated in vivo at Ser137 and Thr210 in mitosis; DNA damage prevents phosphorylation at these sites (14).

  1. Nigg, E.A. (1998) Curr. Opin. Cell Biol. 10, 776-783.
  2. Toyoshima-Morimoto, F. et al. (2002) EMBO Rep. 3, 341-348.
  3. Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-220.
  4. Peter, M. et al. (2002) EMBO Rep. 3, 551-556.
  5. Jackman, M. et al. (2003) Nat. Cell Biol. 5, 143-148.
  6. Nakajima, H. et al. (2003) J. Biol. Chem. 278, 25277-25280.
  7. Sumara, I. et al. (2002) Mol. Cell 9, 515-525.
  8. Hauf, S. et al. (2001) Science 293, 1320-1323.
  9. Peters, J.M. (1999) Exp. Cell Res. 248, 339-349.
  10. Kraft, C. et al. (2003) EMBO J. 22, 6598-6609.
  11. Kotani, S. et al. (1998) Mol. Cell 1, 371-380.
  12. Jang, Y.J. et al. (2002) J Biol Chem 277, 44115-20.
  13. Smits, V.A. et al. (2000) Nat Cell Biol 2, 672-6.
  14. Tsvetkov, L. and Stern, D.F. (2005) Cell Cycle 4, 166-71.

Application References

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Protocols

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and 7,429,487) from Epitomics, Inc.

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

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