Reaktion #358224

Reaktionsgleichung

Hepes

o-NPA

CPT-11

CPT-11

→

SN-38

Reaktionsbedingungen

Aufarbeitung

1. 1
   Sonstigeat 37° C.
2. 2
   Sonstigeup to 20 hours
3. 3
   SonstigeReactions were terminated by the addition of an equal volume

Vorschrift

CE activity was determined by the spectrophotometric method described above for rabbit CE samples using o-NPA as a substrate. In another assay, activation of CPT-11 was determined by incubating samples of hiCE with either 5 μM or 25 μM CPT-11 in a total volume of 200 μl of 50 mM Hepes pH 7.4 at 37° C. for up to 20 hours. Reactions were terminated by the addition of an equal volume of cold acidified methanol, followed by centrifugation at 100,000×g for 30 minutes. The levels of SN-38 produced in the reaction were quantitated by HPLC.

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