Enzyme-catalyzed metal deposition for the enhanced in situ...

G - Physics – 01 – N

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G01N 33/58 (2006.01) C07C 69/017 (2006.01) C07F 9/12 (2006.01) C07H 15/20 (2006.01) C07H 15/24 (2006.01) C07H 15/26 (2006.01)

Patent

CA 2530287

The invention is directed to novel compositions of matter and methods of detecting in situ an immunohistochemical epitope or nucleic acid sequence of interest in a biological sample comprising binding an enzyme-labeled conjugate molecule to the epitope or sequence of interest in the presence of a redox- inactive reductive species and a soluble metal ion, thereby facilitating the reduction of the metal ion to a metal atom at or about the point where the enzyme is anchored. Novel phosphate derivatives of reducing agents are described that when exposed to a phosphatase are activated to their reducing form, thereby reducing metal ions to insoluble metal.

L'invention concerne de nouvelles compositions d'une substance et des méthodes de détection in situ d'un épitope immunohistochimique ou d'une séquence d'acides nucléiques à étudier dans un échantillon biologique consistant à fixer une molécule de conjugué marquée par des enzymes sur l'épitope ou la séquence à étudier en présence d'une espèce réductrice redox inactif et d'un ion métallique soluble, ce qui permet de faciliter la réduction de l'ion métallique en un atome métallique sur ou près du point où l'enzyme est ancrée. L'invention concerne également de nouveaux dérivés du phosphate d'agents de réduction qui, lorsqu'ils sont exposés à une phosphatase sont activés sous leur forme de réduction, réduisant ainsi les ions métalliques en métal insoluble.

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