Homogenization of Atlantic Salmon (S. Salar) Muscle
Instrument: Precellys® 24
Sample: Atlantic salmon (Salmo Salar) muscle
Protocol:
- Add 300mg of frozen salmon muscle to the 2 ml tissue homogenizing kit.
- Add 1 ml extraction buffer (100mM sodium acetate in 0.2% Triton X-100, pH 5.5)*
- Run the Precellys at 5000 rpm, 2 x 20 sec with a 10 sec break.
- Centrifuge at 16,000x g for 30 minutes
*In this instance, the lab was looking to measure cathepsin B. You may substitute a more appropriate buffer for your application, if necessary.
Results:
Salmon muscle tissue was fully and thoroughly homogenized by the Precellys 24, allowing downstream fluorometric detection of Cathepsin activity from the prepared muscle homogenate supernatants.
a) Frozen salmon muscle. b) Muscle prepared for homogenization. c) Muscle post-homogenization. d) Muscle homogenates after centrifugation.
Total cathepsin activity in muscle homogenates from salmon slaughtered without crowding stress (NS), short-term crowding stress (SS) or long-term crowding stress (LS).
