Protocol | Tissue Processing | Protease Inhibitor Additions | ||
---|---|---|---|---|
Tissue Homogenization Buffer | Microsome Resuspension Buffer | |||
A | Scraping | None | 1 mM EDTA | |
B | Scraping | 0.1 mM PMSF | 1 mM EDTA | |
5 mg trypsin inhibitor/g wet weight | 0.1 mM PMSF | |||
C | Mincing | 0.1 mM PMSF | 1 mM EDTA | |
5 mg trypsin inhibitor/g wet weight | 0.1 mM PMSF | |||
D | Scraping | 1 mM EDTA | 1 mM EDTA | |
0.25 mM PMSF | 0.1 mM PMSF | |||
0.1 mM dithiothreitol | 5 μg/ml leupeptin | |||
E | Scraping | 1 mM EDTA | 1 mM EDTA | |
0.1 mM PMSF | 0.1 mM PMSF | |||
1× protease inhibitor mixturea | 1× protease inhibitor mixturea | |||
F | Mincing | 1 mM EDTA | 1 mM EDTA | |
0.1 mM PMSF | 0.1 mM PMSF | |||
5 mg trypsin inhibitor/g wet weight | 0.1 mg/ml trypsin inhibitor | |||
|
| 5× protease inhibitor mixtureb | 5× protease inhibitor mixtureb |