公开(公告)号：US06183978B2

公开(公告)日：2001-02-06

申请号：US09148782

申请日：1998-09-04

Applicant: Irena Bronstein ,  Corinne E. M. Olesen ,  John C. Voyta ,  Yu-Xin Yan

Inventor： Irena Bronstein ,  Corinne E. M. Olesen ,  John C. Voyta ,  Yu-Xin Yan

IPC: C12Q166

CPC classification number: C12Q1/66

Abstract: An assay for the presence of luciferase in a biological sample offers heightened sensitivity, signal intensity and persistence. The assay is sensitive down to 50 fg luciferase. The biological sample is combined with essential ingredients luciferin, ADP, myokinase and Mg++. The myokinase converts ADP to ATP, necessary for the luciferase reaction, and AMP, which retards the reaction kinetics. The resulting assay exhibits a persistent glow emission which makes it adaptable to automation.

Abstract translation: 荧光素酶在生物样品中的存在检测提供了更高的灵敏度，信号强度和持续性。 该测定法灵敏度低至50μg萤光素酶。 生物样品与必需成分荧光素，ADP，肌激酶和Mg ++组合。 肌动蛋白酶将ADP转化为ATP，这是荧光素酶反应所必需的，而AMP则延缓了反应动力学。 所得测定显示持续的辉光发射，使其适应于自动化。

公开(公告)号：US06586196B1

公开(公告)日：2003-07-01

申请号：US09459982

申请日：1999-12-14

Applicant: Irena Bronstein ,  Christopher Martin ,  Corinne Olesen ,  John Voyta ,  Yu-xin Yan

Inventor： Irena Bronstein ,  Christopher Martin ,  Corinne Olesen ,  John Voyta ,  Yu-xin Yan

IPC: C12Q142

CPC classification number: C12Q1/6897 ,  C12Q1/00 ,  C12Q1/34 ,  C12Q1/42 ,  C12Q1/66 ,  G01N2333/916 ,  G01N2333/924 ,  G01N2333/95

Abstract: The present invention discloses multiple enzyme assays which measure the activity of at least one endogenous enzyme in a single aliquot and a method of measuring the activity of multiple enzymes in an aliquot of a cell extract, wherein at least one of the enzymes is an endogenous enzyme. In one embodiment of the invention the activity of a first enzyme is quantified by measuring the light signal produced by degradation of a first enzyme substrate by the first enzyme and the activity of the second enzyme is quantified by measuring the light signal produced by the degradation of a second substrate. In the method of the present invention, both quantifications are performed on the same aliquot of cell extract. Different embodiments of the present invention provide for the detection of more than one endogenous enzyme and for the detection of at least one reporter enzyme and at least one endogenous enzyme. The present invention also discloses kits for detecting the activity of multiple enzymes.

Abstract translation: 本发明公开了测量单一等分试样中至少一种内源性酶的活性的多种酶测定法和测量细胞提取物等分试样中多种酶的活性的方法，其中至少一种酶是内源性酶 。 在本发明的一个实施方案中，通过测量由第一酶降解第一酶底物产生的光信号来定量第一酶的活性，并且通过测量由第一酶降解产生的光信号来量化第二酶的活性 第二基板。 在本发明的方法中，两种定量在相同等分的细胞提取物上进行。 本发明的不同实施方案提供了多于一种内源性酶的检测和用于检测至少一种报道酶和至少一种内源性酶。 本发明还公开了用于检测多种酶的活性的试剂盒。