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检测方法 Colorimetric 描述 概述 The K-LISA™ Akt Activity Kit is a rapid, sensitive, ELISA-based activity assay suitable for measuring the kinase activity of purified or partially purified Akt preparations, in vitro Akt inhibitor screening, and for assessing the regulation of Akt in cell signaling. The assay utilizes a biotinylated peptide substrate (GRPRTSSFAEG) that is phosphorylated on the second serine by Akt1, Akt2, Akt3, SGK (Serum Glucocorticoid Kinase), and MSK1. Biotinylated Akt substrate and sample containing Akt are incubated in the presence of ATP in wells of a streptavidin-coated 96-well plate, which allows for phosphorylation and substrate capture in a single step. 产品目录编号 CBA019 品牌系列 Calbiochem®
应用数据 
Near-confluent MCF-7 cells were stimulated with IGF-1 (100 ng/ml) or IGF-1 (100 ng/ml) and 17β-estradiol (500 nM) for 30 min at 37°C in a CO2 incubator. For inhibition of Akt, cells were first pre-incubated at 37°C in a CO2 incubator for 15 min in the presence of cell-permeable Akt Inhibitor II (Cat. No. 124008) followed by stimulation with IGF-1 (100 ng/ml) for 30 min at 37°C. Cell lysates were prepared using 0.5 ml PhosphoSafe™ Extraction Buffer per 10 cm cell culture dish, per the standard protocol. Cell lysates were either used immediately or stored at -70°C until use. Equal amounts of total protein (1.5 mg) were immunoprecipitated and the activity was determined as outlined in protocol B under the Detailed Protocol section.参考 参考 Liau, S.S., et al. 2006. J. Gastrointest. Surg. 10, 1254.
Hill, M.M. et al. 2002. Pharmacol. Ther. 93, 243.
Shin, I. et al. 2002. Nat. Med. 8, 1136.
Zheng, et al. 2002. J. Biol. Chem. 42, 39379.
Brazil, D.P. et al. 2001. Trends Biochem. Sci. 26, 657.
Medema, R.H., et al. 2000. Nature 404, 782.
Datta, S.R., et al. 1999. Genes Dev. 13, 2905.
Franke, F.T., et al. 1997. Cell 88, 435.
Alessi, D.R., et al. 1996. EMBO J. 15, 6541.产品信息 部件 检测方法 Colorimetric 声明 *Sold under license of U.S. patent 6,441,140 Manufactured by Cell Signaling 形式 96 Tests 格式 96-well plate 试剂盒包含 Streptavidin-Coated 96-Well Plate, Akt Substrate, Biotinylated, Phosphoserine Detection Antibody, Antibody Diluent, Kinase Assay Buffer, 5X ATP/MgCl₂ Mix, 100X Inhibitor, Kinase Stop Solution, ELISA 20X Plate Wash Concentrate, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol. Sold under license of U.S. patent 6,441,140. Manufactured by Cell Signaling. 生物信息 化验范围 6-100 mU/well 化验时间 3 h 样品类型 Cell lysates, tissue extracts, purified enzyme 理化信息 灵敏度 0.05 mUnits 产品使用声明 预定用途 The K-LISA™ Akt Activity Kit is suitable for measuring the kinase activity of purified or partially purified Akt preparations, in vitro Akt inhibitor screening, and for assessing the regulation of Akt in cell signaling. 储存和货运信息 货运代码 Dry Ice Only
毒性 Multiple Toxicity Values, refer to MSDS
有害材料注意事项： Due to the nature of the Hazardous Materials in this shipment, additional shipping chargesmay be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.
储存 -20°C
存储条件 Upon arrival store the Streptavidin-Coated 96-Well Plate at 4°C and the remaining components of the kit at -20°C. 无需冷冻 No
补充信息 试剂盒包含 Streptavidin-Coated 96-Well Plate, Akt Substrate, Biotinylated, Phosphoserine Detection Antibody, Antibody Diluent, Kinase Assay Buffer, 5X ATP/MgCl₂ Mix, 100X Inhibitor, Kinase Stop Solution, ELISA 20X Plate Wash Concentrate, HRP-Antibody Conjugate, TMB Substrate, ELISA Stop Solution, Plate Sealers, and a user protocol. Sold under license of U.S. patent 6,441,140. Manufactured by Cell Signaling.
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