Phospho-eIF4EBP1 (Thr46) Antibody / Eukaryotic translation initiation factor 4E binding protein 1

Western blot analysis of 4EBP1/EIF4EBP1/2/3 (Phospho-T46/T46/T32) using anti-Phospho-eIF4EBP1 (Thr46) antibody. Electrophoresis was performed on a 12% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human HEL whole cell lysates, Lane 2: human SH-SY5Y whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human MCF-7 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-4EBP1/EIF4EBP1/2/3 (Phospho-T46/T46/T32) antibody at 1:500 overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using an ECL Plus Western Blotting Substrate. A specific band was detected for 4EBP1/EIF4EBP1/2/3 (Phospho-T46/T46/T32) at approximately 18 kDa. The expected molecular weight of 4EBP1/EIF4EBP1/2/3 (Phospho-T46/T46/T32) is at 18 kDa.