I'm trying to use ACK lysis buffer to lyse RBCs from whole blood samples after an initial lysing with H2O (restoring isotonicity with 10% NaCl). My last step is using RIPA buffer + protease inhibitors to lyse the WBCs and use the protein lysate for western blotting.
I am concerned with keeping the nuetrophils from activating before I lyse them at the end with RIPA, so I'd like to keep all my spins and reagents cold. I can't find any information about whether this might be an issue for the ACK lysis buffer step - would using cold lysis buffer make it any less effective?
Thanks for the help!
Andrew Kirkham
If you reduce the temperature, you need to increase the time. By how much, you'll have to determine experimentally.
0 votes
0 thanks
- Badges
- Science method
More Madeline Lewis's questions See All
Similar questions and discussions