Anti mouse igg hrp antibody

Cells were collected after stimulated with 100 μM FAC or 100 μM DFO for 24 h. Cellular proteins were extracted by RIPA lysis buffer containing protease and phosphatase inhibitors. SDS-PAGE was used to separate the proteins. After running process, gels were transferred to PVDF membranes and immersed in primary antibodies. The next day, membranes were incubated with secondary antibodies and be visualized by chemiluminescence detection kit (Beyotime). Slc25a28 antibody (ab90170, 1:100) was from Abcam. antibodies specific for SLC25A37/Mitoferrin1 (26469-1-AP, 1:100) and Glut1 (66290-1-Ig, 1:100) were purchased from Proteintech. Anti-phospho-AMPK (Thr172) antibody (#2535S, 1:100), Anti-AMPKα Antibody (#2532, 1:100), anti-p70-S6K (9202S, 1:100), anti-phospo-p70-S6K (Thr389) (9234S, 1:100), anti-Hexokinase 2 (2867S, 1:100), anti-phospho-4EBP1 (Thr70) (13396, 1:100) and anti-4EBP1 (9644s, 1:100) were from Cell Signaling Technology. Anti-PCNA (2586S, 1:100) was from Cell Signaling Technology. The anti-GAPDH antibody (BM1623, 1:1000), anti-β-actin antibody (BM0627, 1:1000), anti-α-tubulin antibody (BM1452, 1:1000), anti-rabbit IgG-HRP antibody (BA1054, 1:5000), and anti-mouse IgG-HRP antibody (BA1050, 1:5000) were purchased from Boster Biological Technology (Wuhan, China).

Cells were collected for protein extraction after treatment with 5 mM NaB for 24 h followed by 25 ng/ml IL-6 for 30 min. RIPA lysis buffer containing protease and phosphatase inhibitors was used to extract cellular proteins. Proteins were separated by SDS-PAGE and then transferred to PVDF membranes and probed with specific primary antibodies and secondary antibodies. An enhanced chemiluminescence detection kit (Beyotime) was used to visualize the bands. Antibodies specific for gp80 (sc-373780, 1:100) and gp130 (sc-376280, 1:100) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies specific for STAT3 (#9139, 1:1000), JAK2 (#3230, 1:1000), p-STAT3 (#9145, 1:1000), p-JAK2 (#3771, 1:1000), and TRAF5 (#41658, 1:1000) were obtained from CST (Beverly, MA, USA). The anti-GAPDH antibody (BM1623, 1:1000), anti-β-actin antibody (BM0627, 1:1000), anti-α-tubulin antibody (BM1452, 1:1000), anti-rabbit IgG-HRP antibody (BA1054, 1:5000), and anti-mouse IgG-HRP antibody (BA1050, 1:5000) were purchased from Boster Biological Technology (Wuhan, China).