Kit for determining glycylproline dipeptide aminopeptidase

A kind of technology of glycylproline dipeptide amino, kit

Inactive Publication Date: 2016-08-17
ANHUI IPROCOM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a test kit for measuring glycylproline dipeptide aminopeptidase in order to overcome the defects of unstable and low measurement accuracy of the existing technology

Method used

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  • Kit for determining glycylproline dipeptide aminopeptidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0057] Reagent R1:

[0058] Tris buffer 215 mmol / L

[0059] Diglycine 8.5 g / L

[0060] Sodium azide 1.1 g / L

[0061] Its solvent is purified water.

[0062] Reagent R2:

[0063] Glycyl-L-proline p-nitroaniline 200 mmol / L

[0064] Ethylene glycol 40mL / L

[0065] Bovine Serum Albumin 7.0 g / L

[0066] Sodium azide 1.1 g / L

[0067] Its solvent is purified water.

Embodiment 2

[0069] The kit of the present invention includes reagent R1 and reagent R2 two-liquid components independent of each other, wherein

[0070] Reagent R1:

[0071] Tris buffer 120 mmol / L

[0072] Diglycine 14.0 g / L

[0073] Sodium azide 1.5 g / L

[0074] Its solvent is purified water.

[0075] Reagent R2:

[0076] Glycyl-L-proline p-nitroaniline 110 mmol / L

[0077] Ethylene glycol 65mL / L

[0078] Bovine serum albumin 3 g / L

[0079] Sodium azide 0.6 g / L

[0080] Its solvent is purified water.

Embodiment 3

[0082] Kit preparation and method of use

[0083] 1. Prepare the reagent according to the content of the following components:

[0084] Reagent R1:

[0085] Tris buffer 215 mmol / L

[0086] Diglycine 8.5 g / L

[0087] Sodium azide 1.1 g / L

[0088] Its solvent is purified water.

[0089] Reagent R2:

[0090] Glycyl-L-proline p-nitroaniline 200 mmol / L

[0091] Ethylene glycol 40mL / L

[0092] Bovine Serum Albumin 7.0 g / L

[0093] Sodium azide 1.1 g / L

[0094] Its solvent is purified water.

[0095] 2. Parameter setting of automatic biochemical analyzer

[0096] (a) Detection temperature: 37°C;

[0097] (b) Detection wavelength: 405nm, sub-wavelength 505nm;

[0098] (c) Reaction time: 8 minutes, among them, the incubation time is 5 minutes, measure and read the absorbance A1 immediately after adding the reagent R2, read the absorbance A2 after 3 minutes, and calculate the change of absorbance ΔA = A2-A1;

[0099] (d) Reaction direction: positive reaction.

[0100] 3....

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Abstract

The invention discloses a kit for determining glycylproline dipeptide aminopeptidase. The kit comprises a liquid reagent R1 and a liquid reagent R2 which are mutually independent. The reagent R1 comprises a 120-310 mmol / L Tris buffer solution, 3.0-14.0 g / L glycylglycine and 0.6-1.5 g / L sodium azide. The solvent of the reagent R1 is purified water. The reagent R2 comprises 110-300 mmol / L glycyl-L-proline nitroaniline, 15-65 ml / L ethanediol, 3-12 g / L bovine serum albumin and 0.6-1.5 g / L sodium azide. The solvent of the reagent R2 is purified water. The kit has the advantages of high accuracy and the like.

Description

technical field [0001] The invention relates to the technical fields of medicine and biochemistry, in particular to a kit for measuring glycylproline dipeptide aminopeptidase. Background technique [0002] Glycylproline dipeptide aminopeptidase (GPDA), which can specifically hydrolyze the peptide bond formed by proline at the end of the peptide chain, uses artificially synthesized X-prolyl-p-nitroaniline as the base When using substances to detect the enzyme activity, it was found that the enzyme activity was the strongest when using glycylproline p-nitroaniline as the substrate, K M The value is the smallest, so this enzyme is called glycylproline dipeptide aminopeptidase or glycylproline p-nitroanilinase, and because it can also hydrolyze glycylprolyl-β-naphthylamine, Therefore, it is also called glycylprolyl-β-naphthylamine enzyme. [0003] GPDA is distributed in the liver, kidney, connective tissue, and salivary glands. Because GPDA can specifically hydrolyze and relea...

Claims

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Application Information

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IPC IPC(8): C12Q1/37
CPCC12Q1/37C12Q2337/12G01N2333/948
Inventor 蔡晓辉庄庆华吴铮徐运
Owner ANHUI IPROCOM BIOTECH CO LTD
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