An Amplex Red-based fluorometric and spectrophotometric assay for L-asparaginase using its natural substrate

Title
An Amplex Red-based fluorometric and spectrophotometric assay for L-asparaginase using its natural substrate
Author(s)
임지석Christos Karamisros[Christos Karamisros]Manfred Konrad[Manfred Konrad]
Keywords
ESCHERICHIA-COLI; OXIDASE; CLONING; MUTANT
Issue Date
201401
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Citation
ANALYTICAL BIOCHEMISTRY, v.445, pp.20 - 22
Abstract
We report on the development of a sensitive real-time assay for monitoring the activity of L-asparaginase that hydrolyzes L-asparagine to L-aspartate and ammonia. In this method, L-aspartate is oxidized by L-aspartate oxidase to iminoaspartate and hydrogen peroxide (H2O2), and in the detection step horseradish peroxidase uses H2O2 to convert the colorless, nonfluorescent reagent Amplex Red to the red-colored and highly fluorescent product resorufin. The assay was validated in both the absorbance and the fluorescence modes. We show that, due to its high sensitivity and substrate selectivity, this assay can be used to measure enzymatic activity in human serum containing L-asparaginase. (C) 2013 Elsevier Inc. All rights reserved.
URI
http://hdl.handle.net/YU.REPOSITORY/33450http://dx.doi.org/10.1016/j.ab.2013.09.028
ISSN
0003-2697
Appears in Collections:
공과대학 > 기계공학부 > Articles
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