Hello all! I have one question about preparing samples for the western blot.. usually I use LDS-sample buffer with b-mercaptoethanol and heat samples at 70 degrees during 10 minutes. But last time when I finished heating I suddenly noticed that the temperature was higher, maybe about 95 degrees. the heating time was the same - 10 minutes..
I wonder what I will see on the western blot in this case? will that heating destroy my samples and I will see an empty wells after staining? I want to check IKK, p-IKK, JNK and p-JNK.
thank you in advance!