What will I actually solubilize if I treat cells with a lysis buffer
that contains 1% tx100 as detergent?
More detailed: Grew hepatoma cells on dishes, washed them with PBS,
froze them in -80, scraped frozen layer into lysis buffer, pipetted up
and down a few times, flash froze in nitrogen, thawed in ultrasound
bath, spun down, I am using the supernatant and kept the pellet. Then
ran westerns with the supernatant.
This is a modified procedure where I used to treat adherent cells
directly with that buffer and the cytoskeleoton and the nuclei remained
on the dish.
I assume that I might get something off the membrane and then just have
What will I get if I potter or ultraturrax the scraped off cells?
Thus, I am looking for some more detailed insights into detergent
fractionation of cells. Any insights you would like to share?