Fluoro NAD

$520.00$2,495.00

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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).

Additional information

Kit Size

100, 500

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

fluorescent_nad_nadh

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

fluorescent_nadh_assay

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

Readable Documents

Title Name
Protocol Protocol.pdf
Datasheet Datasheet.pdf
msds msds.pdf

Kit contents and storage

Part# Reagent Temperature
Part# 6021 Enzyme Mix -20°C
Part# 4018 NADH Detection Reagent -20°C
Part# 7013 NADH Standard -20°C
Part# 3044 NAD/NADH Reaction Buffer 2-8°C
Part# 3045 NAD/NADH Lysis Buffer 2-8°C
Part# 3046 NAD Extraction Buffer 2-8°C
Part# 3047 NADH Extraction Buffer 2-8°C
Part# 3051 NADH Standard Diluent 2-8°C

 

 

References

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

 

Citations

Title File Link Author(s) Journal Year; Edition:Pages
Ethanol disrupts chondrification of the neurocranial cartilages in medaka embryos without affecting aldehyde dehydrogenase 1A2 (Aldh1A2) promoter methylation http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W89-4WXBMB3-1&_user=10&_coverDate=11%2F30%2F2009&_alid=1168344568&_rdoc=4&_fmt=high&_orig=search&_cdi=6649&_docanchor=&view=c&_ct=36&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=912928380dd200a430e7ed4488289345 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 http://aem.asm.org/cgi/content/abstract/AEM.03034-10v1 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 http://aem.asm.org/content/77/9/2905.short 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 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0032062 Schaue D, Ratikan JA, Iwamoto KS PLoS ONE