I've started a brief foray into Western blots. My PI and I have prepared our samples from eye tissue of zebrafish. When we performed the acetone precipitation steps (which we did three times), a dense grey cloud formed in the middle of the sample, leaving our supernatant somewhat convoluted each time we spun it down.
After a Coomassie stain, we saw that whatever that stuff is in the eye tissue was causing the protein to be a thick streak rather than neat little bands. We were able to get a few bands following transfer to a membrane. Is there an extra step of some kind we should be using that is specific to eye tissue?
We obtained exellent results in zebrafihs eyes following the procedure described in J Anat 2010 Sep;217(3):214-22
Thank you for the reference! We tried the protocol in the paper you listed and got the same results that we did with another protocol. I was wondering if you modified their protocol in any way?
Thanks again!
- Badges
- Science method