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泛素-AMC是通過將7-氨基-4-甲基香豆素(AMC)連接到成熟泛素的C末端而制備的,這一過程會(huì)猝滅AMC的固有熒光。當(dāng)與識(shí)別泛素的蛋白酶(例如USP2或UCHL3)孵育時(shí),AMC被釋放出來,熒光強(qiáng)度的增加可以通過在460 nm(激發(fā)波長380 nm)處進(jìn)行測量。LifeSensors的泛素-AMC具有高純度,從而提高了信噪比。該蛋白沒有額外的標(biāo)簽。
艾美捷[干冰]泛素-AMC:
貨號(hào):SI-0220-0050
純度:通過HPLC檢測,純度大于95%
分子量:通過質(zhì)譜檢測為8722 Da
物理狀態(tài):凍干粉
數(shù)量:50 ug
溶解性:在DMSO或0.05%乙酸中溶解度大于1 mg/mL
儲(chǔ)存條件:干粉-20°C保存;溶液-80°C保存,避免反復(fù)凍融。
應(yīng)用:用于測定泛素和泛素樣水解酶的活性和特異性。適用于去泛素化酶活性調(diào)節(jié)劑的高通量篩選。
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文獻(xiàn)參考:
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2) Mason, D.E., Ek, J., Peters, E.C. and Harris J.L. Substrate profiling of deubiquitin hydrolases with a positional scanning library and mass spectrometry. Biochemistry 2004, 43: p6535-44
3) Dang, L. C., F. D. Melandri, and R. L. Stein. Kinetic and mechanistic studies on the hydrolysis of ubiquitin C-terminal 7-amido-4-methylcoumarin by deubiquitinating enzymes. Biochemistry 1998 37: p1868-1879.
4) Stein RL, Chen Z, Melandri F. Kinetic studies of isopeptidase T: modulation of peptidase activity by ubiquitin. Biochemistry. 1995 34(39):p12616-23.
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