Homogenization and Solubilization of RASSLs

1. Place 1 ml of homogenization buffer in a round bottom 14 ml tube.

2. Transfer the frozen tissue into the 14 ml tube. Homogenize using a homogenizer at max speed until tissue is fully ground (about 30-60 seconds).

3. Pipet the homogenate to an eppendorf tube and immediately place it on ice.

4. If the homogenate is too viscous, sonicate for 3-5 seconds.

5. Normalize samples by tissue weight and use approximately 10-50 mg of tissue for solublization.

6. Solublize each sample in an eppendorf tube using the following conditions:

   * 1x SSB
   * 1x Triton-X 100
   * 1x CompleteTM Cocktail (Protease Inhibitor, Boehringer #1697498) 

Example:

x ml of homogenate (~10-50 mg tissue)