Protocol on How to Solubilize Membrane Proteins with CHAPS

Biochemistry, Molecular Biology, and Cell Biology Protocols >> Solubilization of membrane proteins with CHAPS protocol

1. Wash the cells with ice-cold PBS
2. Resuspend the cells in ice-cold TE buffer (10mM Tris, pH 7.4, 2mM EDTA)
3. Wash 2x in ice-cold TE (spin at 40,000 x g, 8 minutes each time)
4. To 36 mM CHAPS in PBS while stirring in the cold (eg on ice or in the cold room), add the cells that are resuspended in ~1mL TE buffer dropwise
5. Solubilize the proteins by stirring for 30 minutes in the cold room after added the cells have been added dropwise
6. Spin the solubilized sample at 100,000 x g, 10 minutes
7. Use the supernatant, which contains the solubilized proteins, for analysis or further purification.