Western Blot Protocol for PPM1B Antibody (NBP1-04335)

Western Blot:
1. Sample preparation: SDS loading buffer was added to MCF7 cell lysates and recombinant PPM1B protein. The solution was boiled before loading on a 15% SDS-PAGE
2. Blocking: 5% skim milk in PBS-Tween, for 1hr at room temperature, 3 times x 10 min PBS-tween washes
3. Primary antibody: Anti-PPM1B antibody(APP0829) 1:2,000, at 4 degrees C;; overnight, 3 times x 10 min PBS-tween washes
4. Secondary antibody: Goat anti-mouse HRP conjugated 1:1,000, for 1hr at room temperature, 3 times x 10 min PBS-tween washes