Learn More
Gibco™ Collagenase, Type IV, powder
Description
Collagenase is unique among proteases in its ability to degrade the triple-helical native collagen fibrils commonly found in connective tissues such as skin, tendon, blood vessels, and bone. Collagenase disaggregation is suitable for the culture of human tumors, mouse kidney, human adult and fetal brain, and many other tissues including epithelium. Collagenase is relatively gentle, dissociates well at physiological temperature and pH, and requires no mechanical agitation or special equipment.
- Isolated from Clostridium histolyticum
- Packaged as lyophilized, non-sterile powder for research use in cell or tissue dissociation and organ perfusions
- Activity is >160 units/mg
- With low tryptic activity and is well-suited for digestion of islet cells from the pancreas
Cell Culture, Mammalian Cell Culture
Order Info
Shipping Condition: Room Temperature
Specifications
Specifications
| Content And Storage | Storage conditions: 2°C to 8°C. Protect from light. Shipping conditions: Room temperature Shelf life: 24 months from date of manufacture |
| Form | Lyophilized |
| Product Type | Collagenase |
| Sterility | Non-sterile |
| Quantity | 1 g |
| Shipping Condition | Room Temperature |
Frequently Asked Questions (FAQs)
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.
Actually, in a feeder-based culture, dispase (2 mg/mL) should take about 15-25 min to work at 37 degrees C. Two to three minutes' dissociation time would apply to feeder-free cultures. Dispase is a more aggressive enzyme, so it works faster, but that also means that when the PSC clumps are harvested, they are more sensitive to being broken apart by trituration. Once the clumps are harvested, they should be pipetted up and down a few times to break up the clumps to the appropriate size. If the cells are harvested with collagenase type IV, they have to be pipetted more times because the clumps are harder to break up, but this means that there is less likelihood to break up the clumps into pieces that are too small. If the cells are harvested with dispase, they have to be pipetted fewer times, and care has to be taken to ensure that the clumps are not broken too much. Either enzyme is fine to use, and if you have enough experience, you may prefer to use dispase to save time. But for a less experienced user, we recommend using collagenase type IV as it is safer and you are less likely to ruin your culture by over-triturating.
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).
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
By clicking Submit, you acknowledge that you may be contacted by Fisher Scientific in regards to the feedback you have provided in this form. We will not share your information for any other purposes. All contact information provided shall also be maintained in accordance with our Privacy Policy.