Enzyme assay from frozen abdominal krill tissue
Instrument: Precellys® 24 with Cryolys cooling device to have a constant temperature of 4°C within the homogenization chamber using liquid nitrogen.
Sample: Abdominal Krill Tissue (frozen)
Protocol:
- Precellys kits: 013961-1-009 (CKmix).
- Sample: ~100 mg (fresh weight) of frozen abdominal krill tissue.
- Extraction solvents: ice-cold deionized water at a concentration of 100 mg fresh weight (fw) mL–1.
- Run the Precellys 24 at 5000 rpm, 2x15sec, 10s break.
- Enzyme assay: Centrifugation of homogenate, spectrophotometrically analysis of supernatant in a specific buffer, data analysis.
Results:
Time course experiments require a standardized and equably homogenization of biological tissues that guarantee a reliable temporal analysis of data. In particular, enzyme assays require a low temperature during the homogenization process to maintain enzyme activity.
During this study, the Cryolys guaranteed a constant temperature of +4°C within the homogenization chamber and thus avoided an uncontrolled defrosting of the frozen samples.
Oscillatory rhythms of the metabolic key enzyme citrate synthase (CS) in light-dark (LD) cycle and in constant darkness (DD).
