Fluoro NAD™
Fluorescent NAD/NADH Detection Kit - Detection of NAD/NADH in tissue extracts/cell samples

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).
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.

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

FluoroNADNADH


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

FluoroNADNADH


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

FluoroNADNADH


NAD and NADH levels were assayed using the Fluoro NAD/NADH kit. Jurkat cells 6 X105 and K562 cells 7 X105 per tube (n=3).


Citations

Inositol 1,4,5-triphosphate receptors and NAD(P)H mediate Ca2+ signaling required for hypoxic preconditioning of hippocampal neurons - P.E. Bickler, C.S. Fahlman, J. Gray, W. McKleroy - Neuroscience Vol 160, Issue 1, April 2009, pp 51-60


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 (2012), PLoS ONE 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 (for 100 assays)
1. Enzyme Mix ..................................................Part# 6021 - Store at -20C
2. NADH Detection Reagent .............................Part# 4018 - Store at -20C
3. NADH Standard .............................................Part# 7013 - Store at -20C
4. NAD/NADH Reaction Buffer ........................Part# 3044 - Store at +2-8 C
5. NAD/NADH Lysis Buffer ..............................Part# 3045 - Store at +2-8 C
6. NAD Extraction Buffer ..................................Part# 3046 - Store at +2-8 C
7. NADH Extraction Buffer ................................Part# 3047 - Store at +2-8 C
8. NADH Standard Diluent ................................Part# 3051 - Store at +2-8 C

 ProductCatalog #Size*Price
Fluoroscent NAD/NADH Detection KitFLNADH 100-2100$495.00
Fluoroscent NAD/NADH Detection KitFLNADH 100-3500$2,375.00

* Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for volume pricing