Gel Electrophoresis of Lactate Dehydrogenase Using Nitroblue Tetrazolium for Enzyme Visualization

Disc Gel Electrophoresis of Lactate Dehydrogenase Using Nitroblue Tetrazolium for Enzyme Visualization 

Prepare solution A (lower gel buffer) by dissolving 56.75 g Tris buffer in 200 ml water. Adjust the pH to 8.9 using concentrated HCl. Add sufficient water to bring the volume to 250 ml. 

Prepare the upper reservoir buffer by dissolving 6 g Tris buffer and 28.8 g glycine in sufficient distilled water to yield a final volume of 1 liter. Check the pH of the solution. If it is not 8.3, adjust the pH with a small amount of Tris or glycine. 

Prepare the lower reservoir buffer by dissolving 908 g Tris buffer in 2 liters water. Add sufficient concentrated HCl to yield pH 8.9. Add sufficient water to yield a final volume of 4 liters. Recheck the pH and adjust if necessary. 

Prepare solution C (ammonium persulfate) by dissolving 17.5 mg ammonium persulfate in 10.0 ml distilled water. 

---