Figure 3. Purification of rSaCas9 by affinity chromatography
The rSaCas9 apoenzyme was purified via its C-terminal Strep-tag II by
affinity chromatography on a Strep-Tactin matrix. Aliquots of 0.5 µl
(IVT pool, pellet, load, flow through, wash) or 6.5 µl (EF#1 – EF#6)
of the different fractions were separated by SDS–PAGE, and the proteins
were visualized by staining with Coomassie Brilliant Blue. Purified
rSaCas9 migrates with an apparent molecular mass of 130 kDa.