Anti hk2 antibody

Total proteins from cell lysate were loaded onto 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel, separated and electro-blotted onto a nitrocellulose membrane (Pierce, Rockford, IL, USA). After blocking with 5% non-fat milk, membranes were probed using various primary antibodies (Anti-iNOS antibody, Abcam, Cat. No.ab178945, 1:2000; anti-Arg-1 antibody, Thermo Fisher, Cat. No.711765, 1:1000; anti-PPARγ antibody, Abcam, Cat. No.ab45036, 1:2000; anti-HK2 antibody, Abcam, Cat. No.ab209847, 1:1000; anti-GLUT1 antibody, Abcam, Cat. No.ab115730, 1:5000; anti-β-actin antibody, Rayantibody, Cat. No.RM2001, 1:2000) at 4 °C overnight, followed with incubation for 1 h with 1:5000 diluted horseradish peroxidase conjugated secondary antibodies (Beyotime Biotechnology, Shanghai, China) at room temperature. The antigen-antibody complexes were detected by the electrogenerated chemiluminescence detection reagent (Meilunbio, Dalian, China). Relative protein expression levels were visualized using Image J software (National Institutes of Health, Bethesda, MD, USA).