Mouse trail elisa kit

Cell supernatants from the different treatment groups were harvested for ELISA. Mice were anesthetized at different time points after ICH induction, and the brain tissue around the bleeding site was used for ELISA. ELISA was performed in strict accordance with the manufacturer’s instructions. The following ELISA kits were used for detection: Mouse IL-1 beta ELISA Kit (Abcam, UK), Mouse TNF alpha ELISA Kit (Abcam, UK), Mouse Tweak ELISA Kit (Abcam, UK), Mouse Activin A ELISA Kit (Abcam, UK), Mouse TRAIL ELISA Kit (Abcam, UK), and Mouse Persephin (Pspn) ELISA Kit (4A Biotech, China).

In order to understand the role of intrinsic apoptosis in anti-tumor responses of the proposed vaccine, the level of caspase-9 and TRAIL in the tumor microenvironment was measured by caspase ELISA kit (Abcam, Cambridge, MA, USA) and Mouse TRAIL ELISA kit (Abcam, Cambridge, MA, USA), respectively. Briefly, one week after the third vaccination, the tumor tissue was extracted from each group (n = 3) and 100 mg of discarded tissue was homogenized in 0.5 ml lysis buffer (0.1 M Tris–HCl pH = 7.6, and 0.1 M fresh dithiothreitol). After centrifugation at 10,000×g (1 min), an equal amount of supernatant was added to the substrate‐containing reaction buffer (0.1 M dithiothreitol and 5 μl of 4 mM DEVD-p-NA) and incubated for 120 min at 37 °C. Finally, the caspase-9 and TRAIL activity was assessed by the microplate reader (BioTek, 800TS, USA) at an absorbance of 405 nm. Each experiment was repeated in triplicate.