Digitonin stock solution preparation

Prepare the digitonin stock solution as described in the section Digitonin Fractionation of Mitochondria. 

A stock solution is prepared by dissolving digitonin (Calbiochem, Bad Soden, Germany) in DMSO at a concentration of 20 mg/ml. The stock solution is divided into small aliquots and stored at -20°C. 

Prepare a 1.5-2% stock solution of digitonin in isolation buffer (see Introduction) without bovine serum albumin. The detergent is dissolved in isolation buffer brought to the boiling point. Once the digitonin is completely dissolved, place the tube on ice. 

Prepare a series of dilutions of digitonin in 100 /itl aliquots of isolation buffer. The lowest concentration has 0% digitonin, while the highest concentration is that of the undiluted stock solution made in step 1. 

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