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半胱天冬酶活性测定

Caspase activity assays.

作者信息

Niles Andrew L, Moravec Richard A, Riss Terry L

机构信息

Promega Corporation, Madison, WI, USA.

出版信息

Methods Mol Biol. 2008;414:137-50. doi: 10.1007/978-1-59745-339-4_11.

Abstract

Caspase activity assays in multi-well plate formats represent powerful tools for understanding experimental modulation of the apoptotic response. These assays are configured to exploit functional, biochemical, and temporal differences in substrate specificity and selectivity, which are useful in defining the magnitude and mechanism of a compound or treatment effect. New advances in fluorescent and luminescent chemistries now enable single addition "add-mix- measure" determinations of caspase activity directly in the sample plate with unprecedented sensitivity. Unlike other more cumbersome or laborious techniques, caspase activity induction or inhibition measures are quantifiable and definitive. The highlighted techniques in this chapter are cost efficient and allow for the rapid exploration of thousands of combinations and conditions.

摘要

多微孔板形式的半胱天冬酶活性测定是理解细胞凋亡反应实验调控的有力工具。这些测定旨在利用底物特异性和选择性方面的功能、生化及时间差异,这有助于确定化合物或治疗效果的大小及机制。荧光和发光化学领域的新进展使得能够在样品板中直接进行单次添加“加样 - 混合 - 测量”来测定半胱天冬酶活性,灵敏度前所未有的高。与其他更繁琐或费力的技术不同,半胱天冬酶活性的诱导或抑制测量是可量化且明确的。本章重点介绍的技术成本效益高,可快速探索数千种组合和条件。

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