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

Karamitros, C. S.; Lim, J.; Konrad, M.

doi:10.1016/j.ab.2013.09.028

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学術論文

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

MPS-Authors

/persons/resource/persons118944

Karamitros, C. S.
Research Group of Enzyme Biochemistry, MPI for biophysical chemistry, Max Planck Society;

/persons/resource/persons82290

Lim, J.
Research Group of Enzyme Biochemistry, MPI for biophysical chemistry, Max Planck Society;

/persons/resource/persons15362

Konrad, M.
Research Group of Enzyme Biochemistry, MPI for biophysical chemistry, Max Planck Society;

External Resource

http://www.sciencedirect.com/science/article/pii/S0003269713004661/pdfft?md5=6e53f99c21987fc6b206d92600928cbb&pid=1-s2.0-S0003269713004661-main.pdf
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1878116.pdf
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1878116_Suppl_1.doc
(付録資料), 3MB

引用

Karamitros, C. S., Lim, J., & Konrad, M. (2013). An Amplex Red-based fluorometric and spectrophotometric assay for L-asparaginase using its natural substrate. Analytical Biochemistry, 445, 20-23. doi:10.1016/j.ab.2013.09.028.

引用: https://hdl.handle.net/11858/00-001M-0000-0014-CF29-6

要旨

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.