Ac-Pro-Ala-Leu-AMC (Ac-PAL-AMC) Summary
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Citation for Ac-Pro-Ala-Leu-AMC (Ac-PAL-AMC)
R&D Systems personnel manually curate a database that contains references using R&D Systems products.The data collected includes not only links to publications in PubMed,but also provides information about sample types, species, and experimental conditions.
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- H727 cells are inherently resistant to the proteasome inhibitor carfilzomib, yet require proteasome activity for cell survival and growthAuthors: MJ Lee, Z Miller, JE Park, D Bhattarai, W Lee, KB KimSci Rep, 2019;9(1):4089.2019
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S-310 hydrolysis confirmation by LCMS-SRMS-310: 1 uMi20S: 15 nMPA28a: 150 nMReaction: 37 °C for 5 h1:S-310 Ac-PAL-AMC (499.25>211.10)2:Free AMC (176.25>120.00)3:S-310 Ac-PAL-COOH (342.20>211.10)

Ordinate: fluorescent count, Em 345/Ex 445Em band path: 9 nmEx band path: 20 nmHorizontal: time [min]Temp: 37 °CS-310: 5, 25, 50, and 100 uMi20S: 100 ng per assay (100 uL)The linearity of the kinetics was confirmed, and the fluorescence intensity increased in a concentration-dependent manner. But it is considered that the saturated level has not been reached using this condition. Probably it could be a low amount of i20S.Compared with S-320, S-310 has slightly higher level of intrinsic fluorescence.

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我在做5RACE,选用TaKaRa公司的RNA反转录酶(RNaseH-),不知道这个酶好不好,有什么需要注意的。
没有序列识别的功能,他就是简单的催化相邻DNA链的5-P末端和3'-OH末端以磷酸二酯键结合的反应,需ATP作辅酶。 不仅可以催化粘性末端之间或平滑末端之间的DNA的连接,也催化DNA与RNA之间以及少数RNA之间的连接。
反转录酶(Reversetranscripatase)是以RNA为模板指导三磷酸脱氧核苷酸合成互补DNA(cDNA)的酶。哺乳类C型病毒的反转录酶和鼠类B型病毒的反转录酶都是一条多肽链。鸟类RNA病毒的反转录酶则由两上亚基结构。真核生物中也都分离出具有不同结构的反转录酶。

