Goat anti rabbit hrp

LPS (lipopolysaccharide) were purchased from Sigma Aldrich Inc (St. Louis, United States), 4′,6-diamidino-2-phenylindole (DAPI) and DIO were obtained from Coollaber Technology (Beijing, China). Dulbecco's modified eagle medium (DMEM) and Fetal Bovine Serum was purchased from Biological Industries (Kibbutz Beit Haemek, Israel). 4% Paraformaldehyde Fix Solution, reactive oxygen species (ROS) assay kit, NO assay kit and BCA assay kit were obtained from Beyotime Biotech-nology Co., Ltd. (Shanghai, China). Quantitative real time polymerase chain reaction (RT-qPCR) reagents were from Vazyme Biotech Co., Ltd. (Beijng, China). DMSO and MTT were from Sangon BiotechCo., Ltd. (Shanghai, China). Antibodies for p38 MAPK, MK2 and β-actin are mouse-derived monoclonal antibodies purchased from Proteintech Group (Wuhan, China). Antibody for CD81, CD63 and Phospho-p38 MAPK (p-p38 MAPK) are rabbit-derived polyclonal antibody purchased from the affinity biosciences LTD (Cincinnati, USA). Secondary antibodies are goat anti-rabbit HRP (Affinity Biosciences, Cincinnati, USA) and goat anti-mouse HRP (Proteintech Group, Wuhan, China), respectively.

The concentration of extracted protein was measured by BCA method. The protein bands were isolated by SDS-PAGE gel electrophoresis and transferred into PVDF membrane (0.45 µm). At room temperature, the PVDF membrane containing the target protein was immersed in the Blotting-Grade blocker (BIO-RAD, California, USA) for 2 h. After blocking, wash three times with buffer solution (TBST). At 4 ℃, the specific primary antibody was incubated overnight. They were washed with buffer solution (TBST) and then incubated with the second antibody for 4 h at room temperature. CD81, CD63 and Phospho-p38 MAPK antibody are rabbit-derived polyclonal antibody purchased from Affinity Biosciences. p38 MAPK antibody, MK2 antibody and β-actin antibody are mouse-derived monoclonal antibodies purchased from Proteintech Group. Secondary antibodies are goat anti-rabbit HRP (Affinity Biosciences) and goat anti-mouse HRP (Proteintech Group), respectively. After washing with buffer solution for three times, test results were obtained by gel imaging system (BIO-RAD ChemiDoc XRS +). Images and results were quantified with Image J software.