Lysis of cell membrane in cell debris using RIPA lysis buffer to extract membrane-bound proteins from the cell debris
I am planning to use RIPA lysis buffer on cell debris to study membrane-bound protein fraction. Yet, I found that mere vortex of the cell debris in RIPA buffer is not sufficient to release proteins from the cell debris. Therefore, I would like to ask for suggestions. Does the process also require specific treatment such as heating of the sample to a certain degree or freezing, etc?