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Membranes were washed with 0.1% Tween-20 and 1 PBS 3 times and incubated with horseradish-peroxidase-conjugated secondary antibodies (Jackson Laboratories) in 5% non-fat skimmed milk powder (Rgilait) in 0.1% Tween-20 and 1 PBS for 1 h at room temperature and washed 3 times with 0.1% Tween-20 and 1 PBS

doi: 10.1124/jpet.107.130351 16 TanakaKHashizumeMMiharaMYoshidaHSuzukiMMatsumotoY

Basically, Im a mess and at the end of my rope trying to fix everything using traditional approaches