Abstract
This chapter describes the assay of thiamine pyrophosphokinase using anion-exchange paper disks. As the separation of thiamine pyrophosphate (TPP) from an assay mixture by chromatography or electrophoresis on paper is rather time consuming, a simple procedure for the rapid radiometric determination of thiamine pyrophosphokinase activity has been designed. Based on its anionic property, labeled thiamine pyrophosphate that is formed during the reaction is bound to diethylaminoethyl (DEAE)-cellulose anion-exchange paper disks, while [ 14 C] thiamine is completely eluted. The assay for thiamine pyrophosphokinase activity is carried out at 37 °. Thiamine monophosphate that might be formed by cleavage of TPP in crude enzyme preparations is also retained by the paper disks, even though to a much lesser extent. Therefore, the enzyme source must be checked for TPP cleaving activities.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
