Product Pathways - Cell Cycle / Checkpoint
Polo-like Kinase Antibody Sampler Kit #9785
|9785S||1 Kit (3 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||Homology†||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|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key: H=Human, R=Rat, Mk=Monkey, M=Mouse
Western blot analysis of recombinant GST-tagged PLK4 Kinase #7580 and extracts from MOLT4, RL and Ramos cell lines using PLK4 Antibody #3258.
Western blot analysis of GSK-PLK1 fusion protein and extracts from HeLa, COS and PC12 cells, using PLK1 (208G4) Rabbit mAb #4513.
Western blot analysis of extracts from HEL, A549 and A20 cells using PLK3 (D14F12) Rabbit mAb #4896.
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.
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.
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.
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).
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.