I'm trying to overexpress a protein of 230 KDa in HEK293. I have been trying for 3 months but I don't see anything on my western blot...
I use Lipofectamine, or Xtrem Gene ( supposed to be better)
I lyse with SDS2%
I use a 4-12% NuPage gel, and i transfer during 3 hours approximately
I use antibodies tthat have been validated on western before...
Any idea to optimize the protocol?
thank you very much,