Reagent for determining .gamma.-glutamyl transpeptidase activity
AssigneeEiken Kagaku Kabushiki Kaisha
InventorKazuhiko Satoh
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Abstract
A reagent for determination of .gamma.-GTP activity containing L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt as a substrate and a transition metal ion or salt, a method for stabilizing L-.gamma.-glutamyl-3-carboxy-4-nitroanilide using a transition metal ion salt, and a method for determining .gamma.-GTP activity using this reagent. The transition metal ion or salt effectively prevents non-enzymatic decomposition (hydrolysis) of the substrate, thereby making it possible to supply the reagent as a solution.
Claims26
A reagent for determining .gamma.-glutamyl transpeptidase activity in a test sample comprising L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof as a substrate and a transition metal ion or salt.
A reagent for determining .gamma.-glutamyl transpeptidase activity in a test sample comprising:
- component A comprising L-.gamma.-glutamyl-3-carboxy-4nitroanilide or a salt thereof as a substrate, a transition metal ion or salt as a stabilizer and a buffer material, and
- component B comprising an amino acid or peptide as a recipient of a .gamma.-glutamyl group and a buffer material.
In a reagent for determining .gamma.-glutamyl transpeptidase activity in a test sample comprising L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof as a substrate, the improvement wherein said L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof is stabilized by the presence of a transition metal ion or salt.
A method for stabilizing L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof comprising adding a transition metal ion or salt to the L-.gamma.-glutamyl-3-carboxyl-4-nitroanilide or a salt thereof.
A method for determining .gamma.-glutamyl transpeptidase activity in a test sample comprising:
- mixing a test sample with component A and component B in any order, wherein said component A comprises L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof as a substrate, a transition metal ion or salt as a stabilizer and a buffer material and wherein component B comprises an amino acid or peptide as a recipient of a .gamma.-glutamyl group and a buffer material,
- measuring a rate of increase of 3-carboxy-4-nitroaniline released from the substrate, and
- determining the .gamma.-glutamyl transpeptidase activity in the test sample based on the rate of increase of 3-carboxy-4-nitroaniline.
A method for determining .gamma.-glutamyl transpeptidase activity in a test sample using L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof as a substrate, said method comprising the steps of:
- (a) mixing a test sample with component A and component B in any order, wherein said component A comprises L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof as a substrate and a buffer material and wherein component B comprises an amino acid or peptide as a recipient of a .gamma.-glutamyl group and a buffer material,
- (b) measuring a rate of increase of 3-carboxy-4-nitroaniline released from the substrate, and
- (c) determining the .gamma.-glutamyl transpeptidase activity in the test sample based on the rate of increase of 3-carboxy-4-nitroaniline,
- wherein said L-.gamma.-glutamyl-3-carboxy-4-nitroanilide or a salt thereof in step (a) is stabilized by the presence of a transition metal ion or salt.
Dependencies are inferred from the claim text.