We used the pervanadate boost method [11 (link)] to increase identification of tyrosine phosphorylated peptides with the isobaric TMT labelling experiments. The pervanadate solution was prepared by adding 10 μl of 0.1 M sodium orthovanadate to 10 μl of 0.2 M hydrogen peroxide (diluted 50 × from a 30% stock). The solution was then incubated at room temperature for 15 min and was added to the PBMCs in PBS. A part of PBMCs from healthy donors were treated with 0.1 mM pervanadate solution in phosphate-buffered saline (pH 7.4) at 37 ℃ for 10 min.
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