Klapperich Lab:Collagen Solution
Protocol for making 0.05% w/v collagen dispersion in acetic acid.
- 5 mL of 0.05M acetic acid
- 2.5 mg of collagen (from human placenta)
- 50 ml beaker
- Ice bath
Making 0.05 M solution of acetic acid
Note: This solution has a shelf life of one week.
- Fill sterilized bottle with 5 mL of Millipore water.
- Add 14.3 µL acetic acid.
0.5% w/v Dispersion of Collagen in Acetic Acid
Note: Keep collagen mixture at below 20°C.
- Add 2.5 mg of collagen to a 10 ml sterilized beaker. (0.05 % w/v)
- Add 5 ml of 0.05 M acetic acid to the beaker.
- Mix solution with homogenizer at full speed for approximately 5 minutes, or until no large fibrils or chunks are apparent. Make sure that tip has been washed thoroughly (ethanol, rinse with Millipore water)
- Place beaker in ice bath.
- Set the sonicater controller to Amplitude 100% (dial), 20 min, 5 sec on, 10 sec off.
- Place sonicater horn in collar, making sure tip of sonicater does not touch the bottom of the beaker. Press START/STOP on sonicater controller.
- The mixture may be stored in a capped bottle at 4°C for about 4 months.
- Re-blend at 4°C at an amplitude of 60 with the pulser set to 5 sec for 15 minutes to use.
Relevant papers and books
- Pieper JS, Oosterhof A, Dijkstra PJ, Veerkamp JH, and van Kuppevelt TH. Preparation and characterization of porous crosslinked collagenous matrices containing bioavailable chondroitin sulphate. Biomaterials. 1999 May;20(9):847-58.