Enzymatic Activity Assay in Macrophages

TG-macrophages (2x10⁵) obtained according to item 2.6 were incubated with RPMI (negative control), BjV (25 μg/mL), and BthTX-I or BthTX-II (25 μg/mL) for 90 min, at 37°C in a humid atmosphere (5% CO₂) in the presence or absence of LED photobiomodulation. After this period, the cells were centrifuged for 1100 xg for 15 min at 4°C and the SOD, catalase, and peroxidase enzymatic activities were determined by commercial kits from Abcam (Cambridge, UK). Absorbances and fluorescences were determined in BioTek Synergy HT MultiDetection (Winooski, VT) at 450 nm and excitation/emission 535/587 nm and expressed in μm/mL or Pm/mL [21 (link), 22 (link)].