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    Reagents and a method for saturation labelling of proteins
    1.
    发明授权
    Reagents and a method for saturation labelling of proteins 有权
    试剂和蛋白质饱和标记方法

    公开(公告)号：US07511155B2

    公开(公告)日：2009-03-31

    申请号：US10519433

    申请日：2002-07-08

    Applicant: Karen Williams ,  Timothy Stone ,  Adrian Christopher Simmonds ,  Alison Claire Sweet ,  Susan Janet Fowler

    Inventor： Karen Williams ,  Timothy Stone ,  Adrian Christopher Simmonds ,  Alison Claire Sweet ,  Susan Janet Fowler

    IPC: C09B23/02 ,  C09B67/22 ,  G01N33/68

    CPC classification number: C07K1/13 ,  C09B23/02 ,  C09B67/0034 ,  G01N33/6815

    Abstract: A matched set of fluorescent dyes is provided, wherein each dye of the set is capable of covalent attachment to a protein and wherein each of the dyes has a molecular structure and a charge that is matched one with the other, such that relative electrophoretic mobility of a protein labeled with one dye of the set is the same as the electrophoretic mobility of the protein labeled with a different dye of the set. The matched set comprises at least two different fluorescent dyes of formula: wherein n is 1, 2, or 3; Z1 and Z2 independently represent the carbon atoms necessary to complete a phenyl or naphthyl ring system; one of groups R1 and R2 is a target bonding group; remaining group R1 or R2 is selected from —(CH2)4—W or —(CH2)r—H; group R3 is hydrogen, except when either R1 or R2 is —(CH2)r—H, in which case R3 is W; and W is selected from sulphonic acid and sulphonate. The invention also provides a method for saturation labeling of a protein with a fluorescent dye so as to label all available target amino acid, suitably cysteine, residues in the protein, thereby giving a single population of labeled protein molecules.

    Abstract translation: 提供了一组匹配的荧光染料，其中该组的每种染料能够与蛋白质共价连接，并且其中每种染料具有彼此匹配的分子结构和电荷，使得相对电泳迁移率 用该组中的一种染料标记的蛋白质与用该组不同染料标记的蛋白质的电泳迁移率相同。 匹配组包含至少两种下式的不同荧光染料：其中n为1,2或3; Z 1和Z 2独立地表示完成苯基或萘基环系所需的碳原子; R 1和R 2中的一个是目标键合基团; 剩余的基团R1或R2选自 - （CH2）4-W或 - （CH2）r-H; R 3是氢，除了当R 1或R 2是 - （CH 2）r-H时，其中R 3是W; W选自磺酸和磺酸盐。 本发明还提供了用荧光染料饱和标记蛋白质的方法，以便标记蛋白质中的所有可用的靶氨基酸，适当的半胱氨酸残基，由此得到单个标记的蛋白质分子群。

    Reagents and a method for saturation labelling of proteins
    5.
    发明申请
    Reagents and a method for saturation labelling of proteins 有权
    试剂和蛋白质饱和标记方法

    公开(公告)号：US20050233462A1

    公开(公告)日：2005-10-20

    申请号：US10519433

    申请日：2002-07-08

    Applicant: Karen Williams ,  Timothy Stone ,  Adrian Simmonds ,  Alison Sweet ,  Susan Fowler

    Inventor： Karen Williams ,  Timothy Stone ,  Adrian Simmonds ,  Alison Sweet ,  Susan Fowler

    IPC: G01N33/58 ,  C07K1/13 ,  C09B23/00 ,  C09B23/02 ,  C09B67/22 ,  G01N21/78 ,  G01N33/68 ,  G01N33/00 ,  C07D43/02

    CPC classification number: C07K1/13 ,  C09B23/02 ,  C09B67/0034 ,  G01N33/6815

    Abstract: A matched set of fluorescent dyes is provided, wherein each dye of the set is capable of covalent attachment to a protein and wherein each of the dyes has a molecular structure and a charge that is matched one with the other, such that relative electrophoretic mobility of a protein labelled with one dye of the set is the same as the electrophoretic mobility of the protein labelled with a different dye of the set. The matched set comprises at least two different fluorescent dyes of formula: wherein n is 1, 2, or 3; Z1 and Z2 independently represent the carbon atoms necessary to complete a phenyl or naphthyl ring system; one of groups R1 and R2 is a target bonding group; remaining group R1 or R2 is selected from —(CH2)4—W or —(CH2)r—H; group R3 is hydrogen, except when either R1 or R2 is —(CH2)r—H, in which case R3 is W; and W is selected from sulphonic acid and sulphonate. The invention also provides a method for saturation labelling of a protein with a fluorescent dye so as to label all available target amino acid, suitably cysteine, residues in the protein, thereby giving a single population of labelled protein molecules.

    Abstract translation: 提供了一组匹配的荧光染料，其中该组的每种染料能够与蛋白质共价连接，并且其中每种染料具有彼此匹配的分子结构和电荷，使得相对电泳迁移率 用该组中的一种染料标记的蛋白质与用该组不同染料标记的蛋白质的电泳迁移率相同。 匹配组包含至少两种下式的不同荧光染料：其中n为1,2或3; Z 1和Z 2独立地表示完成苯基或萘基环系所需的碳原子; 基团R 1和R 2之一是目标键合基团; 剩余的基团R 1或R 2选自 - （CH 2 CH 2）4 -W或 - （CH H 2 H; 基团R 3是氢，除了当R 1或R 2 2是 - （CH 2 2） > r H，在这种情况下R 3为W; W选自磺酸和磺酸盐。 本发明还提供了用荧光染料饱和标记蛋白质的方法，以便标记蛋白质中的所有可用的靶氨基酸，适当的半胱氨酸残基，由此得到单个标记的蛋白质分子群。

    Enhancement of polynucleotide hybridization
    8.
    发明授权
    Enhancement of polynucleotide hybridization 失效
    增强多核苷酸杂交

    公开(公告)号：US5512436A

    公开(公告)日：1996-04-30

    申请号：US108578

    申请日：1993-09-02

    Applicant: Timothy Stone

    Inventor： Timothy Stone

    IPC: C12N15/00 ,  C12Q1/68 ,  G01N1/18

    CPC classification number: C12Q1/6832

    Abstract: Hybridization buffers, for hybridizing complementary polynucleotides, contain polyvinyl alcohol (MW 1000-20000) and/or polystyrene sulphonic acid (e.g. MW 60000-80000) as a rate enhancer, generally at a concentration of 1-10%. Dextran sulphate, polyethylene glycol and cationic detergents may be additionally present. The method is useful when one of the two complementary polynucleotides is immobilised, or is in in situ hybridizations.

    Abstract translation: PCT No.PCT / EP92 / 01479 371日期1993年9月2日 102（e）日期1993年9月2日PCT提交1992年6月30日PCT公布。 出版物WO93 / 01311 日本1993年1月21日。用于杂交互补多核苷酸的杂交缓冲液含有作为速率增强剂的聚乙烯醇（MW 1000-20000）和/或聚苯乙烯磺酸（例如MW 60000-80000），通常浓度为1-10 ％。 可以另外存在硫酸葡聚糖，聚乙二醇和阳离子洗涤剂。 当两个互补多核苷酸之一被固定化或原位杂交时，该方法是有用的。

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