I have homogenized rat plantar skin in lysis buffer and centrifuged at 13 500rpm for 50 minutes. I noticed that there seemed to be 2 phases in the supernatant so recentrifuged it. there are now definatley 2 phases - a white layer AT THE TOP and a clear (majority). I am doing this to probe for NGF by western blot. which phase should I take? what are they? could the top layer be salts?
Adam B Shapiro
The top layer may be lipids. Skim it off and run it separately to see if there is any protein in it.
0 votes
0 thanks
Samantha Locke
Thanks Adam, I came to the same conclusion earlier! out of interest have you ever ran western blots on skin homogenate?
0 votes
0 thanks
Adam B Shapiro
No, I've never done that, but I have seen a white layer on top of sucrose gradients of cell lysates. I skimmed it off and threw it away.
0 votes
0 thanks
- Badges
- Science method
More Samantha Locke's questions See All
Similar questions and discussions