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    Composition and method for measuring thallium influx and efflux
    61.
    发明授权
    Composition and method for measuring thallium influx and efflux 有权
    测量铊流入和流出的组成和方法

    公开(公告)号：US09476892B2

    公开(公告)日：2016-10-25

    申请号：US14521346

    申请日：2014-10-22

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Daniel Beacham ,  Kyle Gee

    IPC: G01N33/53 ,  G01N33/84 ,  G01N33/68 ,  C12Q1/02

    CPC classification number: G01N33/84 ,  C12Q1/02 ,  G01N33/6872 ,  G01N2333/705

    Abstract: The present invention relates to methods for detecting the activity of an ion channel in a cell. The methods comprise providing a loading buffer solution to a cell that has an ion channel. The loading buffer comprises at least one thallium indicator (e.g., an environmentally sensitive, luminescent dye) and a physiological concentration of chloride ions. The methods further comprise providing a stimulus buffer to the cell, wherein the stimulus buffer comprises thallium (e.g., thallium ions). Providing the stimulus buffer causes thallium influx into the cell through the ion channel. After providing the stimulus buffer, the luminescence (e.g., fluorescence) of the dye in the cell is detected. The luminescence of the dye can change in the presence or absence of thallium. The methods may be used to measure influx or efflux of thallium through an ion channel.

    Abstract translation: 本发明涉及用于检测细胞中离子通道的活性的方法。 所述方法包括向具有离子通道的电池提供加载缓冲溶液。 加载缓冲液包含至少一种铊指示剂（例如环境敏感的发光染料）和氯离子的生理浓度。 所述方法还包括向细胞提供刺激缓冲液，其中刺激缓冲液包含铊（例如铊离子）。 提供刺激缓冲液使铊通过离子通道流入细胞。 在提供刺激缓冲液之后，检测细胞中染料的发光（例如荧光）。 在存在或不存在铊的情况下，染料的发光可以改变。 该方法可用于测量铊通过离子通道的流入或流出。

    Optically-Detectable Enzyme Substrates and Their Method of Use
    62.
    发明申请
    Optically-Detectable Enzyme Substrates and Their Method of Use 审中-公开
    光学检测酶底物及其使用方法

    公开(公告)号：US20160139135A1

    公开(公告)日：2016-05-19

    申请号：US14483392

    申请日：2014-09-11

    Applicant: Life Technologies Corporation

    Inventor： Schuyler CORRY ,  William Downey ,  Brian Filanoski ,  Kyle Gee ,  Lawrence Greenfield ,  James Hirsch ,  Iain Johnson ,  Aleksey Rukavishnikov

    IPC: G01N33/58 ,  C07D501/60

    CPC classification number: C07F5/022 ,  C07D499/00 ,  C07D501/00 ,  C07D501/58 ,  C07D501/60 ,  C07D503/00 ,  C07D519/00 ,  C12Q1/34 ,  C12Q2334/00 ,  G01N33/581 ,  G01N2333/986

    Abstract: The present invention relates to compounds that are substrates for an enzyme, and upon reaction with the enzyme provide a detectable response, such as an optically detectable response. In particular, the compounds have utility in detecting the presence of a β-lactamase in a sample. In addition to the compounds, methods are disclosed for analyzing a sample for the presence of a β-lactmase, for example, as an indicator of expression of a nucleic acid sequence including a sequence coding for a β-lactmase. Kits are disclosed that include the disclosed compounds and additional components, for example, cells, antibodies, a β-lactmase or instructions for using the components in an assay.

    Abstract translation: 本发明涉及作为酶的底物的化合物，并且在与酶反应时提供可检测的反应，例如可光学检测的反应。 特别地，这些化合物可用于检测样品中β-内酰胺酶的存在。 除了化合物之外，公开了用于分析样品存在β-乳糖酶的方法，例如，作为包括编码β-乳糖酶的序列的核酸序列的表达的指示剂。 公开了包括所公开的化合物和另外的组分的试剂盒，例如细胞，抗体，β-乳酸酶或在测定中使用组分的说明书。

    Site-specific labeling of affinity tags in fusion proteins
    63.
    发明授权
    Site-specific labeling of affinity tags in fusion proteins 有权
    融合蛋白中亲和标签的位点特异性标记

    公开(公告)号：US09164099B2

    公开(公告)日：2015-10-20

    申请号：US13890137

    申请日：2013-05-08

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Kyle Gee ,  Aleksey Rukavishnikov ,  Courtenay Hart Kerndt ,  Richard Haugland ,  Wai-Yee Leung ,  Wayne Patton ,  Zhenjun Diwu

    IPC: G01N33/533 ,  C07D239/00 ,  C07F5/02 ,  G01N33/58 ,  A61K49/00 ,  C07D311/12 ,  C07D311/18

    CPC classification number: G01N33/52 ,  A61K49/0021 ,  A61K49/0039 ,  A61K49/0041 ,  A61K49/0052 ,  C07D311/12 ,  C07D311/16 ,  C07D311/18 ,  C07F5/022 ,  G01N33/533 ,  G01N33/582

    Abstract: The present invention provides methods and fluorescent compounds that facilitate detecting and labeling of a fusion protein by being capable of selectively binding to an affinity tag. The fluorescent compounds have the general formula A(B)n, wherein A is a fluorophore, B is a binding domain that is a charged chemical moiety, a protein or fragment thereof and n is an integer from 1-6 with the proviso that the protein or fragment thereof not be an antibody or generated from an antibody. The present invention provides specific fluorescent compounds and methods used to detect and label fusion proteins that contain a poly-histidine affinity tag. These compounds have the general formula A(L)m(B)n wherein A is a fluorophore, L is a linker, B is an acetic acid binding domain, m is an integer from 1 to 4 and n is an integer from 1 to 6. The acetic acid groups interact directly with the positively charged histidine residues of the affinity tag to effectively label and detect a fusion protein containing such an affinity tag when present in an acidic or neutral environment.

    Abstract translation: 本发明提供了通过能够选择性地结合亲和标签来促进融合蛋白的检测和标记的方法和荧光化合物。 荧光化合物具有通式A（B）n，其中A是荧光团，B是作为带电化学部分的结合结构域，其蛋白质或片段，n是1-6的整数，条件是 蛋白质或其片段不是抗体或从抗体产生。 本发明提供用于检测和标记含有多组氨酸亲和标签的融合蛋白的特异性荧光化合物和方法。 这些化合物具有通式A（L）m（B）n，其中A是荧光团，L是连接体，B是乙酸结合域，m是1至4的整数，n是1至 乙酸基团与亲和标签的带正电荷的组氨酸残基直接相互作用，以有效地标记和检测当存在于酸性或中性环境中时含有这种亲和标签的融合蛋白。

    Modified Nucleic Acid Binding Cyanine Dyes for the Detection of Reactive Oxygen Species
    65.
    发明申请
    Modified Nucleic Acid Binding Cyanine Dyes for the Detection of Reactive Oxygen Species 有权
    用于检测活性氧物质的改性核酸结合花青染料

    公开(公告)号：US20150079622A1

    公开(公告)日：2015-03-19

    申请号：US14384843

    申请日：2013-02-26

    Applicant: LIFE TECHNOLOGIES CORPORATION

    Inventor： Yi-Zhen Hu ,  Aimei Chen ,  Hee Chol Kang ,  Kyle Gee ,  Bhaskar Mandavilli

    IPC: G01N33/52 ,  C07D417/06

    CPC classification number: G01N33/84 ,  C07D417/06 ,  C09B23/02 ,  C09B23/04 ,  C09B23/06 ,  G01N33/52

    Abstract: Disclosed herein are compounds, compositions, methods, and kits for detecting reactive oxygen species (ROS) by conventional fluorescence microscopy, fluorescence spectroscopy, flow cytometry, and/or high content imaging. The compounds disclosed herein are novel reduced nucleic acid binding cyanine dyes, which dyes are probes for detecting ROS and measuring oxidative stress in cells either in vitro and/or in vivo. Also described herein are processes for preparing novel reduced dyes, i.e., ROS probes, for use in the disclosed compositions, methods and kits.

    Abstract translation: 本文公开了通过常规荧光显微镜，荧光光谱，流式细胞术和/或高含量成像检测活性氧（ROS）的化合物，组合物，方法和试剂盒。 本文公开的化合物是新的还原性核酸结合花青染料，其染料是用于检测ROS的探针，并测定体外和/或体内细胞中的氧化应激。 本文还描述了用于制备用于所公开的组合物，方法和试剂盒的新型还原染料即ROS探针的方法。

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