Hrp conjugated anti rabbit igg secondary antibodies

Liver tissue samples and HepG2 cells were lysed with RIPA lysis buffer (Apply Gene, Beijing, China) containing 1% PMSF. The primary antibodies against CD36 (1:1,000, Abcam, Cambridge, United Kingdom), GAPDH (1:5,000, Proteintech, Chicago, United States), FATP2 (1:1,000, Proteintech, Chicago, United States), and FATP5 (Bioss, Beijing, China) were used. After incubating overnight at 4°C, the corresponding HRP-conjugated anti-rabbit IgG secondary antibodies (1:5,000, Proteintech, Chicago, IL, United States) were used.

Isolated cells were lysed with RIPA lysis buffer supplemented with the protease inhibitor PMSF (Beyotime Biotechnology, China) after washing the cells with PBS. The protein content in the cellular lysate was quantified using the BCA kit (Beyotime Biotechnology, China). The lysate samples were separated on SDS-PAGE after boiling at 100°C for 5 min. The proteins on the gel were cold transferred to a polyvinylidene difluoride membrane (Millipore, United States) for 2 h. After transfer, membrane blocking was done with 5% milk dissolved in PBST, followed by incubation with the target primary antibody for overnight at 4°C with gentle shaking. The membrane was washed and probed with secondary antibodies (Proteintech, China) for 1 h. The protein signal in the membrane was detected using an ultra-high-sensitivity ECL kit (MedChemExpress, United States). Antibodies: CCR9 (Abcam, ab32556), HMGCS1 (Proteintech, 17643-1-AP), MSMO1 (Immunoway, YT7443), HMGCR (HUABIO ET1702-41), MVD (Proteintech, 15331-1-AP), ACTIN (HUABIO, ET1702-52), GAPDH (ABclonal, AC002), HRP-conjugated anti-mouse IgG secondary antibodies (Proteintech, SA00001-1), HRP-conjugated anti-rabbit IgG secondary antibodies (Proteintech, SA00001-2).