Mitochondrial membrane potential assay kit c2006

Anti-α-tubulin (YM3035) and anti-β-tubulin (YM3139) were purchased from ImmunoWay Biotechnology (TX, USA); anti-FASN (SC-48357), anti-NIX (SC-166332), and anti-SREBP1 (SC-13551) were purchased from Santa Cruz Biotech (TX, USA); anti-BNIP3 (D121876) and anti-ACLY (D221957) were purchased from Sangon Biotech(Shanghai, China); anti-ACACA (21923-1-AP) and anti-β-actin (66009-1-Ig) were purchased from Proteintech Group (IL, USA); anti-LC3 II/I (D3U4C) and anti-caspase-3 (D3R6Y) were purchased from Cell Signaling Technology (MA, USA); anti-VDAC1 (AB14734) was purchased from Abcam (MA, USA); D, L-Sulforaphane (574215) was purchased from Sigma-Aldrich (MO, USA); CCCP (C2759) was purchased from Sigma-Aldrich (MO, USA); Baf-A1 (S1413) and PS-341 (S1013) were purchased from Selleck (Shanghai, China); Cell Mitochondria Isolation Kit (C3601), ROS Assay Kit (S0033), and Mitochondrial Membrane Potential Assay Kit (C2006) were purchased from Beyotime Biotechnology (Shanghai, China).

JC-1 staining combined with flow cytometry was employed to determine the mitochondrial membrane potential of mouse hippocampal neurons. In the normal state of mitochondrial membrane potential, JC-1 enters the mitochondrial membrane to form aggregates that are capable of emitting red fluorescence. On the other hand, when the mitochondrial membrane potential decreases, these aggregates break down into monomeric forms capable of emitting green fluorescence. In the present study, the mitochondrial membrane potential assay kit (C2006, Beyotime, Shanghai, China) was used according to the kit instructions for determining the mitochondrial membrane potential of neurons. In brief, neurons were treated with JC-1 (10 µg/mL) for 45 min in the dark and then analyzed using the BD Accuri C6 flow cytometer (BD Bioscience, San Jose, CA, USA).