SBE 13 HCl

Catalog No.S7720

SBE 13 HCl Chemical Structure

Molecular Weight(MW): 479.4

SBE 13 HCl is a potent and selective PLK1 inhibitor with IC50 of 200 pM, >4000-fold selectivity over Aurora A kinase, Plk2 and Plk3.

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Description SBE 13 HCl is a potent and selective PLK1 inhibitor with IC50 of 200 pM, >4000-fold selectivity over Aurora A kinase, Plk2 and Plk3.
Targets
PLK1 [1]
()
200 pM
In vitro

SBE 13 decreases cell proliferation in various cancer cell lines, and causes a G2/M arrest followed by apoptosis. [1] In primary cells, SBE 13 does not impair cell cycle and thus proliferation of primary cells. [2] SBE13 in combination with Enzastaurin displays a synergistic reduction of cell proliferation and enhanced apoptosis induction in HCT116(p53-/-) cells. [3]

Protocol

Kinase Assay:[1]
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Kinase assays:

To assay Plk1 and Aurora A kinase activity, cells are lysed after 13 hrs release in the presence of SBE13 after double thymidine block, and kinases are immunoprecipitated from lysates using antibodies as described. In brief, for each immunoprecipitation 800 µg of total protein were incubated with 1.5 µg Plk1 antibody cocktail, 3 µg Plk2 antibody, 3 µg Plk3 antibody, or 5 µg Aurora A antibody, respectively, for 2 hrs at 4°C on a rotator. Immunoprecipitated protein is collected using Protein G Agarose beads. The Plk1, Plk2 and Plk3 immunoprecipitates are incubated with 1 µg casein and with 1 µCi of [γ32-P]ATP for 30 min at 37°C in kinase buffer. The Aurora A immunoprecipitates are incubated with 0.5 µl Histone and with 1 µCi of [γ32-P]ATP for 60 min at room temperature in kinase buffer. Products from the kinase assays are fractionated on 10% Bis-Tris-polyacrylamide gels, and the phosphorylated substrate is visualized by autoradiography after an exposure of 12 to 36 hrs. An equal amount of immunoprecipitates is subjected to western blot analysis to confirm equal loading of Plk1, Plk2, Plk3 or Aurora A protein in kinase reactions. Immunoprecipitated Plk1 after 13 hrs release in the presence of SBE13 is assayed after de-phosphorylation using λ protein phosphatase and compared to kinase activity of endogenous immunoprecipitated Plk1. Activity of Plk1 kinase with and wiiiuithout de-phosphorylation is compared and the ratio between de-phosphorylated and “normal” endogenous immunoprecipitated Plk1 kinase activity is calculated.
Cell Research:sup>[1]
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  • Cell lines: HeLa, A431, HCT-15, HT-29, MCF-7, U2OS, LN-229, SKW 6.4, PC-3, Det-562, SK-BR-3, SK-OV-3 and A549nb cells
  • Concentrations: 100 μM
  • Incubation Time: 72 hours
  • Method: Cells are treated with SBE13 one day after subculturing. Control cells are incubated with normal culture medium. Concentrations of SBE13 ranged from 1 nM–100 µM. The growth rate of 1 x 105 cells per 6-well is determined by counting cells at 24, 48 and 72 hours after treatment. Cell culture studies are performed in triplicate for each time point.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 95 mg/mL (198.16 mM)
Water Insoluble
Ethanol Insoluble

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 479.4
Formula

C24H28Cl2N2O4

CAS No. 1052532-15-6
Storage powder
in solvent
Synonyms N/A

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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID