Pulldown assay HisTag or GSTTag Pulldown assay HisTag

Pull-down assay (His-Tag or GST-Tag) 一. 實驗原理:

Pull-down assay (His-Tag or GST-Tag) 二. 實驗流程: Incubate Load into beads Wash Elute Incubate (target protein and GST/His tag fused protein) protein conc and final volume : 1 -20 μM , 500 μl temp and time : 4˚C for 4 -24 hours Using spin column or beads: Incubate 3 ~ 5 min 後離心 ( 3000 -6000 rpm) 去掉上清液 Wash 5 min 後離心 ( 3000 -6000 rpm) 去掉上清液 重複wash 3 -5次 (volume : 500 μl) 使用Elution buffer 將target protein洗下來 Volume : 120μl Analysis 使用SDS-PAGE or western blot 分析結果