2.3. Heat stability study
The Cry1Ab/Ac protein was dissolved in 20 mM tris–HCl and 5 mM EDTA buffer
at a concentration of 0.1 mg mL1 in 1.5 mL microcentrifuge tubes. They were
tested for stability at temperature of 100 C for periods of 10, 30, and 60 min. The
assay was terminated by placing the sample tubes on ice, and adding SDS-sample
buffer (50 mM tris–Cl, 8% sucrose, 2% SDS, with 5% 2-mercaptoethanol, and 0.02%
bromophenol blue). Control samples of a 0 min incubation of the protein (kept at
4 C) were also prepared (Hérouet et al., 2005). The resulting proteins were analyzed
by SDS–PAGE and Western Blot.