Klapperich Lab:Collagen Solution

Overview
Protocol for making 0.05% w/v collagen dispersion in acetic acid.

Reagents

 * 5 mL of 0.05M acetic acid
 * 2.5 mg of collagen (from human placenta)

Equipment

 * Homogenizer
 * 50 ml beaker
 * Ice bath
 * Ultrasonicator

Making 0.05 M solution of acetic acid
Note: This solution has a shelf life of one week.
 * 1) Fill sterilized bottle with 5 mL of Millipore water.
 * 2) Add 14.3 µL acetic acid.

0.5% w/v Dispersion of Collagen in Acetic Acid
Note: Keep collagen mixture at below 20&deg;C.
 * 1) Add 2.5 mg of collagen to a 10 ml sterilized beaker. (0.05 % w/v)
 * 2) Add 5 ml of 0.05 M acetic acid to the beaker.
 * 3) 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)
 * 4) Place beaker in ice bath.
 * 5) Set the sonicater controller to Amplitude 100% (dial), 20 min, 5 sec on, 10 sec off.
 * 6) Place sonicater horn in collar, making sure tip of sonicater does not touch the bottom of the beaker. Press START/STOP on sonicater controller.

Contact

 * Jessica
 * Eben