Solubilizing Membrane Proteins with Triton X-100 Protocol

Biochemistry, Molecular Biology, and Cell Biology Protocols >> Solubilizing membrane proteins with Triton X-100 protocol

Solutions required :
10mM Tris, 2mM EDTA (TE buffer)

2% Triton in PBS (make stock solution of 20% Triton X-100 by weighing 2 g Triton X-100, adding PBS up to 10 mL, rocking or mixing gently overnight; store stock solution in the fridge at 4 degrees C)

1. Resuspend cells in TE buffer
2. Wash 2x in TE (spin at 40,000 x g, 8 minutes each time)
3. To 2% Triton in PBS while stirring, add the cells dropwise resuspended in ~1mL TE
4. Solubilize for 30 minutes in the cold room after added cells dropwise
5. Spin at 100,000 x g, 10 minutes
6. Take the supernatant