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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 oxidized 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

 

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
The following kits are available:
Catalog 
 
Size
 
 Price (Eur)
FLNADH 100-2 100 Tests €375

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.

 

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).