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The antibody was purified by affinity chromatography, and conjugated with Alexa Fluor® 488 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 488.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by immunofluorescent intracellular staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is 5 µl per million cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm. ** Alexa Fluor® is a registered trademark of Molecular Probes, Inc. Alexa Fluor® dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
Nocodazole-treated (37°C, overnight) Hela cells were fixed and permeabilized with 70% cold ethanol, then stained with 2A3 Alexa Fluor® 488 vs. Propidium Iodide (DNA content analysis)
Nocodazole-treated (37°C, overnight) (shaded histogram) or non-treated (open histogram) Hela cells were fixed and permeabilized with 70% cold ethanol and then stained with 2A3 Alexa Fluor® 488
PLK-1 (polo-like kinase 1) is a member of te serine/threonine protein kinase family, cdc5/polo subfamily. Highly homologous to polo-like kinase (Drosophila), PLK-1 contains two polo box domains with a predicted molecular weight of 68 kD. This nuclear protein is highly expressed in placenta and colon and has been shown to regulate cdc2/cyclin B through phosphorylation and activation of cdc25c phosphatase. PLK-1 may also be required for cell division; depletion of PLK-1 results in apoptosis. PLK-1 is upregulated by growth stimulating agents and is regulated by cell cycle position (highest in G2/M phase, declining to nearly undetectable levels after mitosis and throughout G1). PLK-1 is modified by phosphorylation (Thr210 is the major phosphorylation site in activated PLK-1 from mitotic cells) and has been shown to interact with nuclear distribution gene C. The 2A3 antibody recognizes human phosphorylated PLK-1 (Thr210) and has been shown to be useful for Western blotting. To increase specificity, it is recommended that the 2A3 antibody be used for Western blotting after immunoprecipitation with the pan-specific PLK-1 3F8 antibody.
Other Names:
Serine/Threonine protein kinase PLK, Polo-like kinase (PLK), Serine-threonine protein kinase 13
Structure:
Serine/Threonine family of protein kinases, cdc5/polo subfamily. Highly homologous to polo-like kinase (Drosophila). Contains two polo box domains. Predicted molecular weight 68 kD
Distribution:
Nuclear protein, highly expressed in placenta and colon
Function:
Regulates cdc2/cyclin B through phosphorylation and activation of cdc25c phosphatase. May be required for cell division. Depletion of PLK-1 results in apoptosis
Regulation:
Upregulated by growth stimulating agents. Regulated by cell cycle position (highest in G2/M phase and declines to nearly undetectable levels after mitosis and throughout G1)
Modification:
Phosphorylation (Thr210 is major phosphorylation site in activated PLK-1 from mitotic cells)
Interaction:
Interacts with nuclear distribution gene C
Antigen References:
1. Hamanaka R, et al. 1994. Cell Growth Differ. 5:249. 2. Lake RJ, et al. 1993. Mol. Cell. Biol. 13:7793. 3. Holtrich U, et al. 1994. P. Natl. Acad. Sci. USA 91:1736.
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This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
Purified anti-PLK-1 Phosphorylated (Thr210)
Panel A. Extracts from untreated Hela cells (Lane 1) or overnight nocodazole-treated Hela cells (Lane 2) were immunoprecipitated with the pan-PLK mAb (clone 3F8), resolved by electrophoresis, transferred to nitrocellulose and probed with mAb 2A3 reactive against Thr210-phosphorylated PLK-1. Proteins were visualized using an HRP goat anti-mouse secondary Ab and a chemiluminescence detection system. Panel B. Extracts from untreated Hela cells (Lane 1) or overnight nocodazole-treated Hela cells (Lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with mAb 2A3 reactive against Thr210-phosphorylated PLK-1. Proteins were visualized using an HRP goat anti-mouse secondary and a chemiluminescence detection system.
Nocodazole-treated (37°C, overnight) Hela cells were fixed and permeabilized with 70% cold ethanol, then stained with 2A3 Alexa Fluor® 488 vs. Propidium Iodide (DNA content analysis)
Nocodazole-treated (37°C, overnight) (shaded histogram) or non-treated (open histogram) Hela cells were fixed and permeabilized with 70% cold ethanol and then stained with 2A3 Alexa Fluor® 488
