1. Add 1.0 ml of deionized water to the Diogenes Reagent that is
contained in the vial. Mix until completely dissolved.
2. Add 9.0 ml of deionized water to the Diogenes Activator that is
contained in the bottle. Mix until completely dissolved.
3. Add the contents in the vial (Diogenes Reagent) to the contents in
the bottle (Diogenes Activator), recap the bottle and shake vigorously
4. The combined solution (10 ml) comprises the Diogenes Complete
Enhancer in its ready-to-use, final working strength, and will yield
up to 100 assays.
1. Add 5X104 - 5X105 cells contained in glucose media into a luminometer
cuvette or microtiter plate.
2. Add 100 μl of Diogenes Complete Enhancer Solution.
3. Add 20 μl of stimulant (PMA, mAb197, fMLP, etc.).
4. Read the results. Time to peak output will depend on the cell and
stimulant used. The response of Diogenes to superoxide is instantaneous.