Hi,

I think Triton X-100 will work fine, I do that to extract a 7-TM lysosomal protein (It is overexpressed though). In my PhD lab, we also used to look at endogenous membrane proteins in mouse kidney and lysed them with a Triton only buffer.

You can lyse the cells by vortexing the extract and ensure good solubilization by incubating the lysates on a wheel at 4°C for 30-60 minutes.

Can you obtain the NP-40 substitute like IGEPAL-CA630 if you want to stick with it?

Good luck

Daniel

If we are not using protease inhibitors what is the best way to homogenize cell lysate?

What cells are you using? You just want to isolate membranes or solubilize them?

NP40 and Triton have very similar properties, the direct replacement for NP40 is IGEPAL CA-630.

Note, however, that the operational criterium of solubilisation is that a protein stays in the supernatant after 1 h at 100000 g. In other words, you'll need an ultracentrifuge, not only a preparative one.