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メルク株式会社
目黒区下目黒1-8-1
153-8927 東京
Japan
TEL： 0120-189-390
Fax: 0120-189-350
E-mail mj_service@merckgroup.com
31 5月 2012
ローディング
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.
| 製品情報 | ||||
|---|---|---|---|---|
| Format | 96-well plate | |||
| Form | 96 Tests | |||
| Detection method | Colorimetric | |||
| Unit definition | One unit is defined to the amount of enzyme required to transfer 1 nmol phosphate per min to substrate at 30° C. | |||
| Components |
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| Sensitivity | 0.05 mUnits | |||
| Assay range | 6-100 mU/well | |||
| Assay time | 3 h | |||
| Sample type | Cell lysates, tissue extracts, purified enzyme | |||
| Kit contains | 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. | |||
| Declaration | *Sold under license of U.S. patent 6,441,140 Manufactured by Cell Signaling | |||
| 保管と送付情報 | |||
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危険物に関する注意： 今回の納品における危険物の特性により、追加の送料が適用される場合があります。サイズによって危険物にかかる追加送料が免除されることもあります。送料に関する詳細はオーダーマネージメントまでお問い合わせください。 |
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| Storage | -20°C | ||
| Ship |
Dry Ice Only
Multiple Toxicity Values, refer to MSDS |
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| データ | |||
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 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 CO<sub>2</sub> incubator. For inhibition of Akt, cells were first pre-incubated at 37°C in a CO<sub>2</sub> 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. |
