Method, substrate and reagents for beta-hexosaminidase A activity detection

A technology of hexosaminidase and hexosaminidase, applied in A, provides a method for the determination of β-hexosaminidase A (in the field of β-Hexosa, which can solve the problem of difficult to achieve the purpose of diagnosis and the inability to truly cause Tay-Sachs disease. Analysis of disease factors, accumulation of diagnostic experience, etc.

A technology of hexosaminidase and hexosaminidase, applied in A, provides a method for the determination of β-hexosaminidase A (in the field of β-Hexosa, which can solve the problem of difficult to achieve the purpose of diagnosis and the inability to truly cause Tay-Sachs disease. Analysis of disease factors, accumulation of diagnostic experience, etc.

CN103484526AInactive Publication Date: 2014-01-01北京中科非凡生物技术有限公司
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  • Method, substrate and reagents for beta-hexosaminidase A activity detection
  • Method, substrate and reagents for beta-hexosaminidase A activity detection
  • Method, substrate and reagents for beta-hexosaminidase A activity detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0062] The β-hexosaminidase A detection reagent of the present embodiment is a single reagent, including

[0063] Disodium hydrogen phosphate / citric acid buffer (pH5.0) 100mmol / L

[0064] Stabilizer 0.1%

[0065] R-β-hexosamine sulfate 0.05mmol / L

[0066] The substrate in the reagent is R-β-hexosamine sulfate, and R is 4-methylumbelliferone.

[0067] The sample of β-hexosaminidase A to be tested is human white blood cells, the total protein dosage is 0.025ug / ul, the volume ratio of protein to reagent is 1 / 19, the sample and reagent are added and mixed to make it react, and the reaction time is 2 Hours, after the reaction is over, stop solution is added to terminate the reaction, and the fluorescence value at excitation wavelength 360nm (Ex=360nm) and emission wavelength 460nm (Em=460nm) is detected with fluorescence detection equipment. The average fluorescence reading value of the reagent blank is 10, and the average fluorescence reading value of normal human white blood c...

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Abstract

The present invention provides a method for beta-hexosaminidase A (EC3.2.1.52) activity detection, and further provides a substrate and reagents for beta-hexosaminidase A activity determination. The method, the substrate and the reagents can be used for analyzing and determining beta-hexosaminidase A activity in a sample (including human body fluid or tissue or cell samples) requiring determination, and are mainly used in the field of clinical laboratory. The determination reagent prepared by using the method has characteristics of convenience, rapidness and high sensitivity, and is easily promoted and applied.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and at the same time belongs to the field of clinical diagnosis and detection of genetic metabolic diseases. Specifically, the present invention provides a method for measuring the enzyme activity of β-hexosaminidase A (β-Hexosaminidase A, EC 3.2.1.52). β-Hexosaminidase A, EC 3.2.1.52) enzyme activity detection substrate, in addition, the invention also provides a reagent for measuring β-hexosaminidase A (β-Hexosaminidase A, EC 3.2.1.52) enzyme activity. Background technique [0002] Lysosome is an organelle wrapped by a unit membrane in human cells. It contains a variety of acid hydrolytic enzymes. More than 60 kinds have been found so far, such as protease, nuclease, glycosidase, lipase, phosphatase, lysozyme and other enzymes. In the cell, lysosomal enzymes control the digestion of various endogenous and exogenous macromolecular substances, such as nucleic acids, proteins, lipids, polysac...

Claims

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Application Information

Patent Timeline
01 Jan 2014
Publication
CN103484526A
IPC
C12Q1/34; G01N21/64
Inventors
孙宏博