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    • 1. 发明申请
    • REAGENTS AND A METHOD FOR SATURATION LABELLING OF PROTEINS
    • 试剂和蛋白质饱和标记方法
    • WO2004005933A1
    • 2004-01-15
    • PCT/GB2002/003142
    • 2002-07-08
    • AMERSHAM BIOSCIENCES UK LIMITEDWILLIAMS, KarenSTONE, TimothySIMMONDS, Adrian, ChristopherSWEET, Alison, ClaireFOWLER, Susan, Janet
    • WILLIAMS, KarenSTONE, TimothySIMMONDS, Adrian, ChristopherSWEET, Alison, ClaireFOWLER, Susan, Janet
    • G01N33/68
    • C07K1/13C09B23/02C09B67/0034G01N33/6815
    • A matched set of fluorescent dyes is provided, wherein each dye of the set is capable ofcovalent 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.
    • 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与另一种相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。