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INTERNATIONAL JOURNAL OF PHARMACEUTICAL RESEARCH

A Step Towards Excellence
Published by : Advanced Scientific Research
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0975-2366
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IJPR 9[3] July - September 2017 Special Issue

July - September 9[3] 2017

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Pros and Cons of some developed methods to measure the intracellular NAD levels

Author: ZAINAB AL-ABADY, ISRAA NAJM ABDULLAH AL-IBADI, NAWAL KHINTEEL JABBAR, ORAAS ADNAN HATEM
Abstract: Nicotinamide adenine dinucleotide (NAD), which known as secret of life is is a cofactor and a key molecule; it plays important rules in signaling pathways, which may contribute to human health known to be regulated upon leukaemia and other cancers. NAD has important role as the only substrate of three consuming enzymes such as CD38/157, PARPs, and sirtuins, these enzymes have been involved in several human diseases, specifically, CD38 which confirms to be the main NAD-regulator enzyme. Thus, NAD is known as a potential target for drug development. Here we draw attention to the importance of finding a sensitive NAD-cycling assay by testing a several NAD-developed methods, the best NAD-cycling assay were selected by make a minor modification on Leorando-cycling assay. In this assay, evaluating the intracellular NAD levels, will also evaluate the effect of NAD-consuming enzymes, or NAD-biosynthesis enzymes. Evaluating intracellular NAD levels may provide a theoretical basis for further drug design. This article is focusing at giving a brief insight into the progress made regarding developed methods to evaluate the intracellular NAD levels, and finally this study provides a sensitive modified assay. The modified ADH cycling method is a convenient method to detect NAD levels, this is a sensitive, and not expensive method can measure NAD levels from (5-60 µM), but not NAD(P) with a short incubation time (30 min), and one catalysing enzyme used rather than two. The ADH-modified method to determine NAD levels in different cell lines may provide novel insights in many disease processes especially in leukemia, were intracellular NAD levels changed in response to the change in the main NAD- regulating enzyme (CD38).
Keyword: NAD, Cycling assays, modified NAD- cycling assay
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