D,L-Sulforaphane N-Acetyl-L-cysteine (SFN-NAC) (CAS 334829-66-2)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
SFN-NAC has better blood-brain barrier permeability than Sulforaphane (SFN).	Treat the cells with 30μM concentration of SFN-NAC for 24 hrs in 6 well plate.

Upstream tips
SFN-NAC has better blood-brain barrier permeability than Sulforaphane (SFN).
Protocol tips
Treat the cells with 30μM concentration of SFN-NAC for 24 hrs in 6 well plate.

Publication protocol

Cells were seeded at a density of 1 × 105 cells/dish in 35-mm cover glass-bottom dishes and cultured for 24 h. Then, SFN-NAC was added to a concentration of 30 uM. The cells were washed three times with PBS after 24 hours, fixed with 4% paraformaldehyde at room temperature for 15 min, and treated with 0.5% Triton X-100 at room temperature for 15 min, after which the solution was discarded. The cells were washed three times with PBS for 3 min each to remove the remaining solution. Next, they were blocked with PBS containing 5% BSA and 0.1% Triton X-100 at room temperature for 1 h. The cells were incubated with the primary antibody overnight at 4°C. The primary antibody was subsequently recovered, and the cells were washed three times for 3 min each with PBS with Tween 20 (PBST). The cells were incubated with the secondary antibody for 1 h at room temperature. The secondary antibody was then discarded, and the cells were washed three times for 3 min each with PBST. A 4′,6-diamidino-2-phenylindole (DAPI)-containing sealer (ZSGB-BIO, Beijing, China) was then added to the center of each dish. Finally, fluorescence images were observed and photographed using a laser confocal microscope (FV1000, Olympus, Tokyo, Japan).

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