1. Add 1 mL Hanks' Balanced Salt Solution (HBSS) with calcium and magnesium directly to 1 g vial of Collagenase. Vortex gently to ensure complete dissolution. Transfer to a clean tube.
2. Determine volume of HBSS (with calcium and magnesium) required to bring collagenase solution to 100 U/µL (1000X stock solution). The activity is lot- specific. Rinse vial with this volume of HBSS (with calcium and magnesium), and combine. Filter sterilize 1000X stock solution with a low protein binding filtration unit.
Example: Assuming the lot you have purchased has an activity of 265 U/mg, this lot will have 265000 Units per mL when you reconstitute collagenase into HBSS (with calcium and magnesium) at 1 g/mL. In order to dilute 265000 U/L to 100000 U/mL (= 100 U/µL), you need to dilute the 1 g/mL enzyme solution 2.65 fold.
Find additional tips, troubleshooting help, and resources within our
Cell Culture Support Center.