alexa Rate assay of N-acetyl-beta-D-hexosaminidase with 4-nitrophenyl N-acetyl-beta-D-glucosaminide as an artificial substrate.
Bioinformatics & Systems Biology

Bioinformatics & Systems Biology

Current Synthetic and Systems Biology

Author(s): Shibata H, Yagi T

Abstract Share this page

Abstract A rapid and accurate rate assay method for N-acetyl-beta-D-hexosaminidase (EC 3.2.1.52, also known as N-acetyl-beta-D-glucosaminidase, or NAGase) using 4-nitrophenyl N-acetyl-beta-D-glucosaminide (NP-GlcNAc) as an artificial substrate was developed using diethylaminoethyl-alpha-cyclodextrin (DEn-CD, where n is the number of diethylaminoethyl groups introduced to alpha-cyclodextrin), as an additive to ionize 4-nitrophenol to yellow-colored 4-nitrophenoxide at pH near 5, where the enzyme acts optimally. A possible recipe for the rate assay of NAGase is as follows. Prepare a stock solution containing 4.8 mmol/l NP-GlcNAc and 1\% DEn-CD (n is preferably near 17) in 0.1 mol/l glycolate buffer, pH 5.50. Introduce the stock solution and a properly diluted sample (urine or other body fluid) to a reaction cell placed in a spectrophotometer at a ratio of 1:1, and monitor the absorbance at 400 or 420 nm. The reaction rate (enzymatic activity) can be conveniently read directly from calibration plots prepared for a given lot of DEn-CD sample, or can be calculated from the rate of the absorbance increase, ionization degree of 4-nitrophenol at pH 5.50, and the millimolar absorbance coefficient of 4-nitrophenoxide in the presence of 0.5\% DEn-CD.
This article was published in Clin Chim Acta and referenced in Current Synthetic and Systems Biology

Relevant Expert PPTs

Relevant Speaker PPTs

Relevant Topics

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri, Food, Aqua and Veterinary Science Journals

Dr. Krish

[email protected]

1-702-714-7001 Extn: 9040

Clinical and Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemical Engineering and Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001 Extn: 9040

Earth & Environmental Sciences

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

General Science and Health care Journals

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics and Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001 Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Informatics Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Material Sciences Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Mathematics and Physics Journals

Jim Willison

mathem[email protected]

1-702-714-7001 Extn: 9042

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001 Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

John Behannon

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001 Extn: 9042

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords