NOS

  

Nitric Oxide Synthase Detection Kit (Cell Permeable)

 

1.  Key Benefits 

 

•  Cell permeable.

•  No wash homogenous assay.

•  Adaptable to High throughput assay platforms .

•  Real time detection of NO activity.

•  Applications – Flow Cytometry, Fluorescence Plate Reader, Fluorescence Microscopy

 

2.  Assay Principle

Diaminofluorescein –2 Diacetate (DAF-2DA) is a non-fluorescent cell permeable reagent that can measure free Nitric Oxide (NO) and nitric oxide synthase (NOS) activity in living cells under physiological conditions. Once inside the cell the diacetate groups on the DFA-2DA reagent arehydrolyzed by cytosolic esterases thus releasing FAD-2 and sequestering the reagent inside the cell. Production of nitric oxide converts the non-fluorescent dye, DAF-2, to its fluorescent triaole derivative, DAF-2T.

 

DAF-2T can be observed by: excitation 488nm and measuring emission at 515nm.

 

Fig. 1. DAF-2DA, the non-fluorescent probe, enters the cells where cytosolic esterases hydrolysis the diacetate groups, releasing the non-fluorescent DAF-2 dye. DAF-2 then reacts with NO and O2 to produce DAF-2T, its triazole fluorescent derivative.

 

Fig. 2. Phase Contrast and fluorescent microscopic images of rat aorta-derived vascular smooth muscular cells loaded with DAF-2DA.

 

Reagent	Catalog #	Size
Nitric Oxide Synthase	NOS200-1	0.125 mg
Nitric Oxide Synthase	NOS200-2	0.25 mg

 

NOS Protocol

 

NOS Datasheet

 

msds.NOS