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Using N,N,N’,N’-tetramethyl-p-phenylenediamine (TMPD) to Assay Cyclooxygenase Activity In Vitro

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Book cover Advanced Protocols in Oxidative Stress II

Part of the book series: Methods in Molecular Biology ((MIMB,volume 594))

Abstract

Prostaglandin endoperoxide synthase (PGH synthase), also known as cyclooxygenase (COX), was identified over 30 years ago and is the key enzyme in the pathway by which arachidonic acid is converted to the range of biologically active lipid mediators known as the prostanoids that participate in numerous physiological processes. The need for the development of new and improved COX inhibitors as potential therapeutics also drives the need for rapid, reliable, and inexpensive assays of COX activity. Colorimetric assays are often the preferred methods of enzyme analysis since they may be readily adapted to simple microplate formats that require relatively inexpensive and widely available instrumentation. The use of N,N,N ¢,N ¢-tetramethyl-p-phenylenediamine (TMPD) in high throughput microplate assays of COX activity could become the approach of choice in the screening of potential therapeutics that inhibit COX activity in vivo. Considering that TMPD is also a potential substrate for most, if not all, heme peroxidases, it is anticipated that this agent could find increasing application in the future.

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Author information

Authors and Affiliations

  1. School of Pharmacy, Murdoch University, Murdoch, WA, Australia

    Nenad Petrovic

  2. Pharmacogenetics and Drug Development Group, University of Sydney, Sydney, NSW, Australia

    Michael Murray (Faculty of Pharmacy)

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  1. Nenad Petrovic
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  2. Michael Murray
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Correspondence to Nenad Petrovic .

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  1. Golden Leaf Rd. 215, Mars Hill, 28754, USA

    Donald Armstrong

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Petrovic, N., Murray, M. (2010). Using N,N,N’,N’-tetramethyl-p-phenylenediamine (TMPD) to Assay Cyclooxygenase Activity In Vitro. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress II. Methods in Molecular Biology, vol 594. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-411-1_9

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  • DOI: https://doi.org/10.1007/978-1-60761-411-1_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-410-4

  • Online ISBN: 978-1-60761-411-1

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