| 1. Anesthesize mouse with 0.02 ml 2.5% Avertin/gram body weight. |
| 2. Make incisions through the skin and the peritoneum and cut out approximately 100-200 mg of the tissues to be analyzed. |
| 3. Immediately rinse the tissue in 1x PBS, blot dry, and weigh. |
| 4. Transfer the tissue into an eppendorf tube and freeze in liquid nitrogen. |
| 5. After all the samples are collected, they may be stored at -70°C or used for immunoprecipitation. |
- 2.5% Avertin (Chem Name = Tribromoethanol, Fluka #90710)
- Dilute 100% Avertin in warm PBS (55°C or higher)
- Filter sterilize, wrap in foil, store frozen aliqouts at -20°C
- Aliqouts good for ~3 weeks at 4°C
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- 100% Avertin
- Dissolve Avertin in tert-amyl alcohol (Sigma #A-1685)
- Wrap in foil, store at 4°C, good for years.
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- *Note: Tribromoethanol is extremely light sensitive. Its degradative products, HBr and dibromylacetaldehyde, are both very toxic.
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