Reagents for Oxidative Burst Assay

Luminol (sodium salt), lucigenin (N, N′-dimethyl-9,9′-biacridinium dinitrate), phobol 12-myristate 13-acetate (PMA), G418, casein (from bovine milk, sodium salt), and 4-aminobenzoic hydrazide (4-ABAH, myeloperoxidase inhibitor-1; Calbiochem, EMD Millipore) were purchased from Sigma-Aldrich (Sigma-Aldrich, MO, USA). L-012 sodium salt was purchased from Wako Chemicals USA (Richmond, VA, USA). Verdiperstat, AZD5904, and hydroxypropyl-β-cyclodextrin (HP-β-CD) were purchased from MedChemExpress (NJ, USA). Anti-CTLA-4 (9D9), αPD-1 (RMP1-14), CD8α (2.43), CD4⁺ (GK1.5), NK1.1 (PR136), and polyclonal rat IgG control were obtained from Bio X cell (NH, USA). Luminol sodium salt was dissolved in sterile phosphate-buffered saline (PBS) to a final concentration of 50 mg/mL or 100 mg/mL and stored at −20°C. L-012 powder was dissolved in sterile double-distilled water (ddH₂0) to a final concentration of 20 mM and stored at −20°C. 4-ABAH was dissolved in sterile ddH₂O heated to 65°C to a final concentration of 2 mg/mL.44 (link) Verdiperstat and AZD5904 were dissolved in sterile DMSO to a final concentration of 10 mM and stored at −20°C; AZD5904—5% DMSO, 15% HP-β-CD w/v, 200 µL intraperitoneal injection daily. Verdiperstat—4% DMSO,15% HP-β-CD w/v, 300 µL intraperitoneal injection daily. Nine grams of casein was dissolved in 100 mL PBS, pH7.2 containing 0.9 mM CaCl₂ and 0.5 mM MgCl₂.45 (link)