Component Amount Stock concentration
Collagenase IV (Invitrogen 20 000 units (typically 200 U/mL in D-MEM Catalog no. 17104-019) 1 mg/mL)
Dissolve 20 000 units of collagenase IV in 100 mL of D-MEM (usually 1 mg/mL). Filter using a 250 mL filter unit. Aliquot in 5-10 mL tubes and store at —20°C until use.
■ Collagenase is isolated from Clostridium histolyticum. Type IV is selected because of low tryptic activity and is recommended for isolation of pancreatic islets.
■ This is a crude product, so expect lot-to-lot variation.
■ EDTA inhibits this enzyme's activity.
■ A unit is defined as the amount of enzyme required to liberate 1 pM of L-leucine equivalents from collagen in 5 h at 37°C at pH 7.5.
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