Extended Data Fig. 4: Protocol used for the detection of myristoylated Trx-h2 in Arabidopsis.
From: Redox-dependent structural switch and CBF activation confer freezing tolerance in plants
Azidomyristate was vacuum-infiltrated into 2-week-old Trx-h2-V5OE/trx-h2 and Trx-h2(G/A)-V5OE/trx-h2 plants incubated at 22 °C for 24 h. Total proteins extracted from these plants were incubated with phosphine-PEG3-biotin. The biotinyl-myristoylated Trx-h2-V5 was immunoprecipitated with anti-V5 antibody and separated by SDS-PAGE on a reducing gel. Then, the biotinyl-myristoylated Trx-h2-V5 was detected by western blot analysis using anti-biotin antibody. The protocol was established by modifying the method used for the detection of myristoyl proteins in cell cultures21.
