Isolation of protein for Western blots from engineered tissues

1.        Make an appropriate volume of 1X Laemmli sample buffer (LSB)

2.        Wash constructs 2X with ice cold PBS

3.        Place 150l of Laemmli sample buffer

4.        Sonicate the cells for 30 seconds

5.        Mix by hand

6.        Sonicate for another 30 seconds

7.        Heat 5 minutes at 100C

8.        Store at -80C or continue on to gel