Lysis and running:
Cells lysed in 1.5X Lammeli buffer +0.15M DTT syringed 10X with 25g needle and then boiled 100 degrees C for 10 minutes.
10ug of sample run out on 4-12% Bis Tris Nupage gel in 1X MOPS buffer.
Western blot was performed by wet transfer of gel onto nitrocellulose membrane 0.22u pore size\n 100V for 2 h 4 degrees.
Blocked in 5% milk PBST for 1 h room temp (1X PBS + 0.1% tween 20)
incubated 24h at 4 degrees in 5% PBST with primary antibody Rabbit polyclonal to CRISPR-Cas9 ab204448) 1:1000.
Washed 3X PBST;
IgG anti rabbit HRP 1:5000 (Jackson Immunoscience);
Washed 3x PBST;
Washed 1X PBS;
Developed using ECL prime kit (GE Healthcare).
Abcam user community
Submitted Feb 14 2019
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