Fluoraldehyde o phthaldialdehyde reagent solution opa

Mitochondrial superoxide was measured with the MitoSOX™ Red mitochondrial superoxide indicator for live cell imagining (Molecular Probes, Thermo Fisher Scientific, Switzerland). SH-SY5Y cells were seeded in a 96-well plate (Huberlab, Switzerland) at a density of 3.5 × 10⁴ cells and cultured as described above. The measurements were done according to the supplier’s datasheet. Briefly, a [5 mM] stock solution of the provided MitoSOX reagent was prepared in DMSO and the stock solution was further diluted to a [5 µM] working solution in Hank’s balanced salt solution with calcium and magnesium (HBSS, Sigma, Switzerland). At DIV8, non-differentiated, differentiated as well as nanoparticle-treated SH-SY5Y cells were incubated with [5 µM] MitoSOX working solution at 37 °C for 50 min after the cells were washed with HBSS. Fluorescence measurements were performed immediately at an emission of 510 nm and excitation of 580 nm with a Synergy H1 multi-mode reader (BioTek, Switzerland). Subsequently, the protein amount of each sample was determined using Fluoraldehyde™ o-phthaldialdehyde reagent solution (OPA, Thermo Fisher Scientific, Switzerland) at an emission of 360 nm and an excitation of 460 nm. The results of the MitoSOX Assay were normalized to 1 mg protein.

bovine serum albumin was purchased to engineer the albumin conjugates. bovine serum albumin (BSA), locust bean gum (LBG, M_w = 66,000 g/mol), and sodium alginate (ALG, medium viscosity, M_w = 120,000 g/mol) were purchased from Sigma Aldrich Ltd. (Hamburg, Germany). The Fluoraldehyde™ o-Phthaldialdehyde Reagent Solution (OPA) was obtained from Thermo Scientific™ (Waltham, MA, USA). Broad-range SDS–PAGE molecular weight standards purchased from VWR (VWR International Ltd., Debrecen, Hungary) were used as molecular mass standards (myosin 200 kDa, β-galactosidase 120 kDa, bovine serum albumin 91 kDa, glutamic dehydrogenase 62 kDa, ovalbumin 47 kDa, carbonic anhydrase 37 kDa, myoglobin 28 kDa, lysozyme 19 kDa, aprotinin 9 kDa). The distilled water was prepared by a laboratory purification system.