Authors
Suaib Luqman, Nusrat Masood, Suchita Srivastava, and Vijaya Dubey
Summary
A precise method has been developed for measuring the activity of Ornithine decarboxylase (ODC) enzyme, a promising target of cancer and infectious diseases using petite quantity of chemicals. The modified spectrophotometric assay has been used to find inhibitors of ODC enzyme. The methodology is based on the cognizance that enzyme transforms L-ornithine hydrochloride (substrate) to yellow colored putrescine (product) soluble in pentanol, the absorbance of which was measured spectrophotometrically. The technique described herein was optimized for evaluating efficacy of phytomolecules as ODC attenuator but could also be used for exploration of novel bioactive compounds as chemopreventive and/or chemotherapeutic agents. A reaction concoction was prepared and hydrophobic withdrawal of yellow colored TNP-Putrescine-TNP complex was recorded at 426nm. Advanced validation and understanding of the factors affecting the enzyme activity i.e. substrate concentration, inhibitor concentration, temperature, pH of the reaction medium and incubation time were also investigated.Further details
The protocol was published on Protocol Exchange on 22 April 2013. To see the entire protocol, click on the source link.Advertisement
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