Fluoro NAD

Product Description

Introduction

The role of pyridine nucleotides (NAD/NADH) metabolism in health, is of continual and increased interest. A growing amount of evidence supports the fact that NAD metabolism regulates important biological effect including life span. NAD, through poly-ADP-ribosyl polymerase (PARP), mono-ADP-ribosyltransferase (ARTs) and recently characterized sirtuin enzymes, exerts potential biological effects. These enzymes modify proteins to regulate their function via ADP-ribosylation or deacetylation and are involved in several pathways including apoptosis, DNA repair, senescence and endocrine signaling. This suggests that either the enzymes or NAD could be an important therapeutical target 1.

Key Benefits

• Detection of NAD/NADH content in cells or tissue extracts.
• Detection of NAD/NADH levels in apoptosis, metabolism, proliferation, DNA repair, senescence, endocrine signaling and life span.
• NAD/NADH detection in Bacterial, fungal and plant cells.
• Fluorescent 96 well Plate Reader Readout (excitation: 530-570nm and emission at 590-600nm).

Assay Principle

The Fluoro NAD/NADH detection kit utilizes a non-fluorescent detection reagent, which is reduced in the presence NADH to produce its fluorescent analog and NAD. NAD is further converted to NADH via an enzyme coupled reaction. The enzyme reaction specifically reacts with NAD/NADH and not with NADP/NADPH
Reaction:

1. NADH + non-fluorescent detection reagent+ electron  →  coupler fluorescent

analog + NAD

2. NAD + enzyme coupled reaction  → NADH (then proceeds to reaction 1).

excitation: 530-570nm and emission at 590-600nm

Figure 1. NADH standard curve titrated in NAD/NADH lysis buffer. Incubation time = 1 hour.

Figure 2. NAD/NADH Assay showing no cross-reactivity with NADPH. Incubation time = 1 hour.

Citations

Ethanol disrupts chondrification of the neurocranial cartilages in medaka embryos without affecting aldehyde dehydrogenase 1A2 (Aldh1A2) promoter methylation - Yuhui Hu, Kristine L. Willett, Ikhlas A. Khan, Brian E. Scheffler and Asok K. Dasmahapatra - Comparitive Biochemistry and Physiology Part C: Toxicology & Pharmacology Vol 150, Issue 4, November 2009, pp 495-502

High titer anaerobic 1-butanol synthesis in Escherichia coli enabled by driving forces - Claire R. Shen, Ethan I. Lan, Yasumasa Dekishima, Antonino Baez, Kwang Myung Cho, and James C. Liao - Appl. Environ. Microbiol March 11th 2011. doi:10.1128/AEM.03034-10

Driving Forces Enable High-Titer Anaerobic 1-Butanol Synthesis in Escherichia coli - Claire R. Shen, Ethan I. Lan, Yasumasa Dekishima, Antonino Baez, Kwang Myung Cho and James C. Liao - App Environ Microbiology Vol 77, No 9, pp 2905-2915, May 2011

Cellular Autofluorescence following Ionizing Radiation - Schaue D, Ratikan JA, Iwamoto KS - PLoS ONE (2012); 7(2): e32062

References

• Anthony A. Sauve NAD+ and Vitamin B3: From metabolism to therapies J. Pharmacol. Exp. Ther. 2007 : jpet.107.120758v1

Kit contents and Long Term storage