Collagenase, Type I, powder, 1 g - FAQs

View additional product information for Collagenase, Type I, powder - FAQs (17018029, 17100017)

3 product FAQs found

Can collagenase be used to dissociate feeder free hESC when cultured on CELLstart medium?

Yes, collagenase can be used. It is recommended to use a 10 mg/ml collagenase IV solution (0.5 ml per 6 cm dish). After 1 minute, remove collagenase and rinse cells. Scrape cells into wash buffer (see product manual for more information), pellet, and re-plate as desired.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

How do I make the 1000X stock (100 U/µL) solution from Collagenase powder?

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.

What reagents do you offer for cell dissociation, and what are the main differences between them?

Please use this selection chart that compares our cell dissociation reagents (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/reagents/trypsin.html).

Find additional tips, troubleshooting help, and resources within ourMammalian Cell Culture Basics Support Center.