.cp_wz table {border-top: твердое тело 1px #ccc; border-left: твердое тело 1px #ccc; } .cp_wz таблица td {border-right: 1px solid #ccc; нижняя граница: сплошной 1px #ccc; padding: 5px 0px 0px 5px;} .cp_wz table th {border-right: 1px solid #ccc; border-bottom: 1px solid #ccc; padding: 5px 0px 0px 5px;} \ n Молекулярный вес: \ n 479,4 SBE 13 HCl - мощный и селективный ингибитор PLK1 с IC50 200 пМ,> 4000-кратная селективность по сравнению с киназой Aurora A, Plk2 и Plk3. \ n Биологическая активность SBE 13 снижает пролиферацию клеток в различных линиях раковых клеток и вызывает остановку G2 / M с последующим апоптозом. В первичных клетках SBE 13 не нарушает клеточный цикл и, следовательно, пролиферацию первичных клеток. SBE13 в комбинации с энзастаурином демонстрирует синергетическое снижение клеточной пролиферации и усиление индукции апоптоза в клетках HCT116 (p53 - / -). Протокол (только для справки) Анализ киназы: [1]
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Kinase assays
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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.
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Клеточный анализ: sup> [1]
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Cell lines
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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
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Concentrations
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100 μM
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Incubation Time
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72 hours
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Method
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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.
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Конверсия различных модельных животных на основе BSA (значение на основе данных из проекта рекомендаций FDA)
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Species
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Baboon
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Dog
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Monkey
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Rabbit
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Guinea pig
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Rat
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Hamster
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Mouse
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Weight (kg)
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12
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10
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3
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1.8
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0.4
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0.15
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0.08
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0.02
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Body Surface Area (m2)
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0.6
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0.5
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0.24
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0.15
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0.05
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0.025
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0.02
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0.007
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Km factor
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20
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20
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12
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12
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8
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6
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5
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3
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Animal A (mg/kg) = Animal B (mg/kg) multiplied by
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Animal B Km
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Animal A Km
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Например, чтобы изменить дозу ресвератрола, используемую для мыши (22,4 мг / кг), на дозу, основанную на BSA для крысы, умножьте 22,4 мг / кг на коэффициент Km для мыши, а затем разделите на коэффициент Km для крыса. Этот расчет дает эквивалентную дозу ресвератрола для крыс 11,2 мг / кг.
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Rat dose (mg/kg) = mouse dose (22.4 mg/kg) ×
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mouse Km(3)
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= 11.2 mg/kg
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rat Km(6)
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Химическая информация
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Molecular Weight (MW)
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479.4
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Formula
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C24H28Cl2N2O4
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CAS No.
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1052532-15-6
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Storage
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3 years -20℃Powder
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6 months-80℃in solvent (DMSO, water, etc.)
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Synonyms
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N/A
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Chemical Name
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N-(4-((6-chloropyridin-3-yl)methoxy)-3-methoxybenzyl)-2-(3,4-dimethoxyphenyl)ethanamine hydrochloride
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Калькулятор молярности Калькулятор разведения Калькулятор молекулярной массы
Группа Продуктов : Клеточный цикл > Ингибитор PLK
