Lactate dehydrogenase cytotoxicity test kit

The levels of TNF-α and IL-1β (Novus Biologicals, Inc.) in pleural effusions and the blood of mice were determined. Anti-CD8 antibody (Abcam), anti-PD1 antibody (EPR20665, Abcam), anti-PDL1 antibody (EPR20529, Abcam), anti-GAPDH antibody (ab8245, Abcam), and anti-β-actin antibody (13E5, CST) were used for antigen detection. Lymphocyte and tumor cell lines were co-cultured in 96-well plates at a ratio of 20:1. The LDH release experiment was then performed using the lactate dehydrogenase cytotoxicity test kit (Beyotime Biotechnology).

All test kits used here were provided by Beyotime Biotechnology Co., Ltd. Lactate dehydrogenase cytotoxicity Test Kit (Cat. No. C0017) was used to detect the release of LDH in R28 cells to determine cytotoxicity in line with the guidelines of supplier. Lipid Oxidation (malondialdehyde, MDA) Detection Kit was performed to quantitatively detect the MDA level of R28 cells (Cat. No. S0131S) strictly as per reagent directions. After OGD/R stimulation, the levels of ROS and SOD in R28 cells with or without gastrodin treatment were detected by ROS detection kit (Cat. No. S0033S) and SOD activity detection kit (Cat. No. S0101M), respectively. Intracellular glutathione peroxidase activity was detected by corresponding assay kit (Cat. No. S0056). Enzyme-linked immunosorbent assay (ELISA) kit was employed to examine the levels of intracellular inflammatory factors TNF-α (Cat. No. PT516), IL-6 (Cat. No. PI328) and IL-1β (Cat. No. PI303).