| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Phospho-PLK1 (Thr210) Rabbit mAb Coated Microwells | 77588 | 96 tests |
|
+4C |
| PLK1 Rabbit Detection mAb | 17378 | 1 ea |
|
+4C |
| HRP Diluent | 13515 | 5.5 ml |
|
+4C |
| TMB Substrate | 7004 | 11 ml |
|
+4C |
| STOP Solution | 7002 | 11 ml |
|
+4C |
| Sealing Tape | 54503 | 2 ea |
|
+4C |
| ELISA Wash Buffer (20X) | 9801 | 25 ml |
|
+4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml |
|
-20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The rapid protocol (RP) PathScan® RP Phospho-PLK1 (Thr210) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PLK1 protein phosphorylated at Thr210 in a reduced assay time of 1.5 hours. Incubation of cell lysates and detection antibody on the coated microwell plate forms a sandwich with phospho-PLK1 (Thr210) in a single step. The plate is then extensively washed and TMB reagent is added for signal development. The magnitude of absorbance for the developed color is proportional to the quantity of phospho-PLK1 (Thr210). Learn more about all of your ELISA kit options here.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
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).
- Nigg, E.A. (1998) Curr Opin Cell Biol 10, 776-83.
- Toyoshima-Morimoto, F. et al. (2002) EMBO Rep 3, 341-8.
- Toyoshima-Morimoto, F. et al. (2001) Nature 410, 215-20.
- Peter, M. et al. (2002) EMBO Rep 3, 551-6.
- Jackman, M. et al. (2003) Nat Cell Biol 5, 143-8.
- Nakajima, H. et al. (2003) J Biol Chem 278, 25277-80.
- Sumara, I. et al. (2002) Mol Cell 9, 515-25.
- Hauf, S. et al. (2001) Science 293, 1320-3.
- Peters, J.M. (1999) Exp. Cell Res. 248, 339-49.
- Kraft, C. et al. (2003) EMBO J 22, 6598-609.
- Kotani, S. et al. (1998) Mol Cell 1, 371-80.
- Jang, Y.J. et al. (2002) J Biol Chem 277, 44115-20.
- Smits, V.A. et al. (2000) Nat Cell Biol 2, 672-6.
- Tsvetkov, L. and Stern, D.F. (2005) Cell Cycle 4, 166-71.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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