Anti rabbit phospho 4e bp1 thr37 46

The phosphorylation levels of mTOR and 4-EBP were measured to determine influences of FM replaced by FSM on protein metabolism in the muscle of large yellow croaker. The detailed procedures were according to the previous study by Chen et al. [23 (link)]. The primary antibodies are anti-rabbit mTOR (1 : 1000, #2972, Cell Signaling Technology; MA, USA), anti-rabbit phospho-mTOR (Ser2448) (1 : 1000, #5536, Cell Signaling Technology; MA, USA), anti-rabbit 4E-BP1 (1 : 1000, #9452, Cell Signaling Technology; MA, USA), anti-rabbit phospho-4E-BP1 (Thr37/46) (1 : 1000, #2855, Cell Signaling Technology; MA, USA), anti-mouse β-actin (1 : 5000, AC004, ABclonal Technology; Wuhan, China), as well as anti-rabbit β-tubulin (1 : 5000, AC008, ABclonal Technology; Wuhan, China), respectively. The secondary antibodies are HRP-conjugated anti-rabbit IgG antibody (1 : 10000, 7074, Cell Signaling Technology; MA, USA) and HRP-conjugated mouse anti-rabbit IgG antibody (1 : 10000, 5127, Cell Signaling Technology; MA, USA). Quantification of protein bands was accomplished by the ImageJ software.