Enzyme activity determinations method characterized by the using of substrates whose fluorescent properties differs those of the converted products
First Claim
1. In the method for demonstrating the presence of an activity of an enzyme by(a) incubating said enzyme with a fluorogenic substrate A which is converted by the enzyme to a product B differing from A in respect to its fluorescent properties and with standardized amounts of remaining substances necessary for the activity, and(b) measuring the change in fluorescence caused by said enzyme, the improvement being that at least one of (1) said substrate A and (2) said product B is provided with a chromophore which is a triplet sensitizer having a triple energy level above the excitation energy level of a lanthanide ion selected from the group consisting of Eu3+, Tb3+, Dy3+ and Sm3+ and which is can chelate said lanthanide ion by means of an oxygen or nitrogen atom in said chromophore, and that B differs from A either by(i) carrying a different chromophore, or(ii) having a different chelating activity.
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Abstract
The method for demonstrating the presence of an activity of an enzyme by
(a) incubating said enzyme with a fluorogenic substrate A which is converted by the enzyme to a product B differing from A in respect to its fluorescent properties, A and/or B carrying a chromophore which is a triplet sensitizer having a triplet energy level above the excitation energy level of a lanthanide ion selected from the group consisting of Eu3+, Tb3+, Dy3+ and Sm3+ and which is capable of chelating said lanthanide ion by means of an oxygen or nitrogen atom in said chromophore, and that B differs from A either by
(i) carrying a different chromophore, or
(ii) having a different chelating ability, and
(b) measuring the change in fluorescence caused by said enzyme.
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Citations
9 Claims
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1. In the method for demonstrating the presence of an activity of an enzyme by
(a) incubating said enzyme with a fluorogenic substrate A which is converted by the enzyme to a product B differing from A in respect to its fluorescent properties and with standardized amounts of remaining substances necessary for the activity, and (b) measuring the change in fluorescence caused by said enzyme, the improvement being that at least one of (1) said substrate A and (2) said product B is provided with a chromophore which is a triplet sensitizer having a triple energy level above the excitation energy level of a lanthanide ion selected from the group consisting of Eu3+, Tb3+, Dy3+ and Sm3+ and which is can chelate said lanthanide ion by means of an oxygen or nitrogen atom in said chromophore, and that B differs from A either by (i) carrying a different chromophore, or (ii) having a different chelating activity.
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