The reaction was incubated at RT for 30 min and fractionated in 5 % non-denaturing polyacrylamide gel, followed by electrophoretic transfer to nylon membrane.
Fifteen micrograms protein was run by SDS / PAGE and transferred to nitrocellulose membranes using a Mini Trans - Blot Electrophoretic Transfer Cell (Bio — Rad, CA, U.S.A.).