NAO staining has been used to measure mitochondrial content in cells. The dye binds specifically to cardiolipin in mitochondria. Neurons were seeded in a 6-well-plate and transfected as detailed above. NAO dye (400 nM, Sigma Aldrich, St. Louis, MO, USA) was added to cells at 14–16 DIVs in mKRB. Cells were stained for 30 min at 37 °C, washed with mKRB, detached mechanically by a soft scraping and suspended in mKRB at the concentration of 100,000 cells/mL. Mitochondrial mass was assessed by flow cytometry using the FACS Canto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA).
Nao dye
Manufactured by Merck Group
Sourced in United States
NAO dye is a fluorescent staining agent used in biological research laboratories. It is a small molecule that binds to nucleic acids, allowing for the visualization and detection of DNA or RNA in various experimental procedures. The core function of NAO dye is to provide a fluorescent signal that can be detected using appropriate imaging equipment, such as fluorescence microscopes or flow cytometers.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Facscanto 2, by BD (1 mentions)
Anti lc3b, by Merck Group (1 mentions)
Alexa fluor 488 conjugated, by Abcam (1 mentions)
Rapamycin rapa, by Selleck Chemicals (1 mentions)
Parp1, by Proteintech (1 mentions)
Myd88, by Merck Group (1 mentions)
Ndufb8, by Proteintech (1 mentions)
Anti sod1, by Proteintech (1 mentions)
Anti beclin 1, by Bioss Antibodies (1 mentions)
P62 sqstm1, by Merck Group (1 mentions)
St2825, by MedChemExpress (1 mentions)
8 ohdg, by Bioss Antibodies (1 mentions)
Uqcrfs1, by Proteintech (1 mentions)
Anti ki67, by Abcam (1 mentions)
3 methyladenine 3 ma, by Selleck Chemicals (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
2 protocols using nao dye
1
Mitochondrial Content Quantification by NAO
2
Oxidative Stress, Autophagy, and Apoptosis
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High‐glucose (25 mM) Dulbecco's modified Eagle's (DMEM‐H), foetal bovine serum (FBS), antibiotics (penicillin and streptomycin), and Lipofectamine2000 were obtained from Thermo Fisher Scientific. Methyl viologen dichloride hydrate (paraquat, PQ), dichloro‐dihydro‐fluorescein diacetate (DCFH‐DA), DAPI, acridine orange (AO), dansylcadaverine (MDC), and NAO dye were purchased from Sigma (MO, USA). Rapamycin (RAPA) and 3‐methyladenine (3‐MA) were purchased from Selleckchem. ST2825 was purchased from MedChemExpress. All molecular biological reagents were obtained from New England Biolabs. Primary antibodies against mTOR, phosphor(P)‐mTOR, P‐p53, Drp1, Tom20, cleaved‐caspase 3, and Bcl‐2 were purchased from Cell Signalling Technology. Anti‐Ki67, phosphorylated histone H2A.X (P‐γH2A.X), CD3, and CD68, and Alexa Fluor® 488‐conjugated or Alexa Fluor® 594‐conjugated second antibody were purchased from Abcam. Anti‐LC3B, SQSTM1/P62, and MyD88 were purchased from Sigma. Anti‐Beclin 1, CDK2, GPX1, 8‐OHdG, OPA1, and Fis1 were purchased from Bioss (Woburn, MA, USA). Anti‐SOD1, p53, PARP1, NDUFB8, and UQCRFS1 were purchased from Proteintech. Secondary antibodies horseradish peroxidase‐conjugated goat anti‐rabbit or mouse IgG was purchased from ZSGB‐Bio.
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