Zinc sulfate znso4

Luteolin ≥98% (LT) was supplied by Baoji Guokang Bio-Technology Co. (Baoji, China). Lipoid^®S100 (LP) l-α-phosphatidylcholine, M.wt = 786.1 g/mol was obtained from Lipoid AG (Ludwigshafen, Germany). Cholesterol (CH) was a gift from the Nile Company for Pharmaceuticals and Chemical Industries (Cairo, Egypt). Streptozotocin (STZ) was obtained from Sigma–Aldrich (St. Louis, MO, USA). Vanadium trichloride, N-1-naphthyl-ethylenediamine, sulfanilamide, sodium nitrate (NaNO₃), and zinc sulfate (ZnSO₄) were purchased from Sigma–Aldrich (Munich, Germany). All other chemicals and reagents used were of analytical grade.

1 × 10⁶ EL-4 T-cells were incubated with a final allicin concentration of 25 µM for 30 min with gentle shaking at 37 °C in the dark. Afterwards, cells were washed and loaded with 1 mL measurement buffer [32] (link) for 30 min, containing 1 µM FluoZin-3AM (Thermo Fisher, Germany) and again gently shaken at 37 °C in the dark. Control cells were not treated with allicin. Cells were washed, resuspended in measurement buffer and incubated at 37 °C for 10 min either left untreated or further supplemented with either N,N,N’,N’-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN, 50 µM) to obtain minimal fluorescence or with a combination of zinc sulfate (ZnSO₄) and pyrithione (100 µM/50 µM) (all Sigma-Aldrich, Germany) to obtain maximal fluorescence. Subsequent flow cytometry measurements were performed using FACSCalibur (BD, Germany). Calculation of intracellular labile zinc was performed as described before [33] (link) using the dissociation constant K_D = 8.9 nM for the FluoZin-3/Zn²⁺ complex [34] (link).

Pyrrole (C₄H₄NH, ≥98%, Sigma Aldrich, Munich, Germany), Zinc sulfate (ZnSO₄, ≥98%, Sigma-Aldrich, Munich, Germany), hydrochloric acid (HCl, ≥37%, Sigma-Aldrich, Munich, Germany), cadmium nitrate (Cd(NO₃)₂, ≥98%, Sigma-Aldrich, Munich, Germany) and sodium hydroxide pellets (NaOH, N98%, Sigma-Aldrich, Munich, Germany), sodium dihydrogen phosphate (NaH₂PO₄.2H₂O, 99.95%, Sigma-Aldrich, Munich, Germany), potassium persulfate (K₂S₂O₈, 99.99%, Sigma-Aldrich, Munich, Germany), silver nitrate (AgNO₃, ≥99%, Sigma-Aldrich, Munich, Germany) and chloroform (CHCl₃, 99%, Sigma-Aldrich, Munich, Germany) were purchased and used. All the reagents used are of AR grade and used as received.