Il 6 elisa kit

EN was purchased from Shanghai CP Guojian Pharmaceutical Co., Ltd. (Shanghai, China). HA (molecular weight 300kDa) was obtained from Freda Biochem Co., Ltd. (Shandong, China). Methacrylic anhydride (MA), 2-Hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (I 2959), N,N′-methylenebisacrylamide (MBA) and Complete Freund’s Adjuvant (CFA) were obtained from SIGMA-ALDRICH (Merk, Darmstadt, Germany). Sulforhodamine B (SRB) was purchased from Yuanye Biotechnology Co., Ltd. (Shanghai, China). Trypan blue was purchased from Beyotime Biotechnology (Shanghai, China). TNF-α ELISA kit was purchased from Solarbio Life Science (Beijing, China) and IL-6 ELISA kit was purchased from MultiSciences (Hangzhou, China). All chemicals for preparation and analysis were obtained from the commercial source and used as received. Ultrapure water (18 MΩ cm; GenPure, Thermo Fisher Scientific, Waltham, MA, USA) was used for all solution preparations.

To measure the circulating TNF-α, IL-6, and IL-1β levels in vivo, blood was removed from the eyes. The samples were stored frozen before sample testing. The levels of TNF-α, IL-6, and IL-1β in the murine serum samples were analyzed using TNF-α ELISA kit (70-EK282HS-96; MULTI SCIENCES), IL-6 ELISA kit (70-EK206HS-96; MULTI SCIENCES), and IL-1β ELISA kit (EK201B/3-96; MULTI SCIENCES) according to the manufacturer’s instructions.

NP cells were trypsinized, dispersed into single-cell suspensions and seeded in six-well plates. Each well contained 5 × 104 cells. Then the culture medium was added with visfatin (PeproTech, 96–130-09-25). The concentration of IL-6 in the cell supernatant was detected using an IL-6 ELISA Kit (MultiSciences, 70-EK106/2-96) and visfatin in the rats plasma collected from the rats blood samples was detected using an visfatin ELISA Kit (Abcam, ab267799), according to the manufacturer’s instructions.