Conjugationally extended hydrazine compositions of the formula (RR
2
)N(H)
n
(NH
2
)
n
, fluorescent hydrazone compositions of the formula (RR
2
)NN═C(R
1
R
2
), methods of the formation of hydrazones from the reaction of conjugationally extended hydrazines with conjugationally extended carbonyls and methods of their use in assays systems are described. Use of these conjugationally extended hydrazine and oxime compositions for direct calorimetric and fluorometric assays wherein a chromophore or the fluorophore is incorporated into the linker that is positioned between a reactive linking moiety and a biotin molecule. More specifically the linker comprises one molecule of a high affinity binding pair such as for example biotin of the biotin/avidin high affinity binding pair, connected to a spacer molecule such as for example a length of polyethyleneglycol followed by a pro-chromophoric, chromophoric, pro-fluorescent or fluorescent moiety connected to an amino-, thiol- or carbohydrate-reactive moiety such as for example succinimidyl, maleimido or aminoxy group respectively, that may covalently link to a biomolecule.
描述了公式(RR2)N(H)n(NH2)n的共轭扩展
肼组合物,公式(RR2)NN═C(R1R2)的荧光
肼酮组合物,以及从共轭扩展
肼与共轭扩展羰基化合物反应形成
肼酮的方法,以及在测定系统中使用它们的方法。利用这些共轭扩展
肼和
肟组合物进行直接量热和荧光测定,其中色团或荧光团被并入连接基团中,该连接基团位于反应性连接基团和
生物素分子之间。更具体地,连接基团包括一分子高亲和性结合对之一,例如
生物素/亲和素高亲和性结合对之一,连接到一个间隔分子,例如一段聚
乙二醇长度,后接一个前色团、色团、前荧光团或荧光团,连接到一个
氨基、
硫醇或
碳水化合物反应性基团,例如琥珀
酰亚胺基、马来
酰亚胺基或
氨氧基团,可以共价连接到
生物分子。