Asante NaTRIUM Green Loading Protocol

  

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Asante NaTRIUM Green Loading Protocol

The following is supplied as a starting point for non-invasive loading of ANG-1 via the acetoxymethyl (AM) ester version.  Due to cell type and other experimental variations, loading conditions will require optimization.

1. Prepare a 1 mM stock solution of ANG-1 (AM) in anhydrous dimethylsulfoxide (DMSO).  
   
   • For a 500 µg vial, dissolve the contents of the vial in 450 µL DMSO.  
   • For a 50 µg vial, use 45 µL DMSO.
2. Divide the stock solution into aliquots that will be consumed per experiment.
   
   • Store the aliquots at -20 ºC, protected from moisture and light.
   • Repeated freezing and thawing of a stock solution typically leads to degradation of the product via hydrolysis of the AM esters
   • For your convenience, we supply ANG-1 (AM) in 50 µg packages.
3. Dilute the stock solution to twice the original volume with a solution of 20% Pluronic F-127 in DMSO.
   
   
4. Disperse the ANG-1/Pluronic F-127 solution into 100 times the volume of serum-free culture medium.
   
   • The final ANG-1 (AM) concentration is 5 µM.  (The concentration of ANG-1 AM may require optimization.)
   • Pluronic F-127 is 0.1% of the final solution.
5. Incubate the cells for one to two hours at room temperature.  (Incubation time and temperature may also require optimization.)
   
   
6. Remove the cell loading medium and wash the cells with serum-free and dye-free medium.
   
   
7. The cells are now loaded with ANG-1 (AM) and ready for your sodium imaging experiments.

Learn more about Asante NaTRIUM Green-1 and Asante NaTRIUM Green-2.

Download the complete ANG Info Packet HERE