Ldh cytotoxicity assay kit c0017

Germacrone (CAS 6902-91-6) was purchased from Weikeqi (China) and dissolved in dimethyl sulfoxide (DMSO) at 20 mM. Lithium chloride (LiCl) (Sigma, St. Louis, MO, USA) and diammonium glycyrrhizinate (DG) (TargetMol, USA) were diluted in DMEM at 1 M and 10 mg/mL concentration, respectively, and filter-sterilized. The following primary antibodies were purchased from commercial resources: rabbit anti-β-Actin pAb from HuanBio (China); mouse anti-PRV-gB mAb (LD-DW-Z0022-2) from lv du (China). The secondary antibodies used in western blotting, including DyLight 800 goat anti-mouse IgG (H+L) (074-1806) and DyLight 800 goat anti-rabbit IgG (H+L) (074-1506), were purchased from KPL (Gaithersburg, MD). Secondary antibody used in confocal microscopy, including goat anti-mouse–FITC (Fluorescein isothiocyanate) (172-1806) was obtained from KPL. The lactate dehydrogenase (LDH) cytotoxicity assay kit (C0017) was purchased from Beyotime (China).

The procedures for C. sakazakii infection of HBMEC were the same as the NO release assay. LDH release was detected using an LDH Cytotoxicity Assay Kit C0017 (Beyotime). The absorbance at 490 nm of samples was measured with a microplate reader (Victor X3; PE, Singapore) to evaluate LDH release after C. sakazakii infection. The percentage of LDH release was calculated as follows: % LDH release = (sample LDH activity-background LDH)/(total LDH activity-background LDH) ×100.