Cells were washed in phosphate-buffered saline (PBS) three times, incubated with 200 nM MitoTracker Deep Red (Beyotime Biotechnology) at 37 °C for 30 min, fixed with 4% paraformaldehyde for 30 min, and permeabilised with 0.1% Triton X-100 for 10 min. After blocking with 1% BSA for 1 h, the cells were incubated with diluted primary antibody (ALDH2 at 1:200 ratio) overnight at 4 °C. After washing, the cells were incubated with the corresponding FITC-conjugated secondary antibodies (1:200, 1 h at 37 °C). The nuclei were stained with 4′,6-diamidino-2-phenylindole for 5 min at room temperature. Immunofluorescence micrographs were captured using Nikon AXR confocal laser microscope.
Mitotracker deep red
Manufactured by Beyotime
Sourced in China
MitoTracker Deep Red is a fluorescent dye that selectively stains mitochondria in live cells. It is a membrane-permeant, red-fluorescent dye that can be used to visualize and track mitochondria in a variety of cell types.
Automatically generated - may contain errors
Lab products found in correlation
Axr confocal laser microscope, by Nikon (1 mentions)
Mrfp gfp lc3 adenovirus, by Hanbio Biotechnology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Lyso tracker red, by Beyotime (1 mentions)
Er tracker red, by Beyotime (1 mentions)
4 protocols using mitotracker deep red
1
Mitochondrial Protein Localization Imaging
2
Mitochondrial Network Analysis in MRC5 Cells
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Cultivate MRC5 cells on coverslips in a 24-well plate, followed by overnight incubation. Upon cell adhesion, treat the cells with lactate, Mdivi-1, and Ulixertinib. Subsequently, stain the cells with the mitochondria-specific fluorescent probe MitoTracker Deep Red (100 nM, purchased from Beyotime) for 30 min. Conduct a 10-min treatment of MRC5 cells with 0.1% Triton X-100, followed by DAPI restaining to label cell nuclei. Finally, fix the coverslips onto glass slides and capture images using confocal microscopy. Perform quantitative analysis of the mitochondrial network using the MiNA plugin in ImageJ.
3
Visualizing Autophagy in OM-MSCs
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OM-MSCs were briefly transfected with GFP-mRFP-LC3 adenovirus (20 multiplicity of infection; Hanbio, Shanghai, China) for 24 h, and Mito-Tracker Deep Red (30 min at 37°C; Beyotime) was used to detect the localization of mitochondria. The nuclei were stained with DAPI (Beyotime). Yellow dots were detected in at least 100 cells in each individual experiment by confocal microscopy after different treatments. Image Pro-Plus software was used to assess the degree of co-location using Manders overlap coefficients.
4
Phytic Acid Interaction with Metal Ions
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Phytic acid was purchased from McLean Biochemical Technology Co., Ltd. (Shanghai, China). Ethylenediamine, NaCl, FeCl3·6H2O, CaCl2, FeCl2, MgCl2·6H2O, HgCl2, MnSO4·H2O, CuSO4·5H2O, ZnCl2, ammonium acetate, potassium permanganate, and potassium dichromate were provided by the Damao Chemical Reagent Plant (Tianjin, China). All amino acids were purchased from Mellon Biotechnology Co., Ltd. (Dalian, China). 3-(4,5-Dimethylthiazole)-2,5-phenyltetrazolium bromide (MTT) and dimethyl sulfoxide (DMSO) were purchased from Sigma (Shanghai, China). ER-Tracker Red, Lyso-Tracker Red, and Mito-Tracker Deep Red were obtained from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Cells were provided by Procell Life Science & Technology Co., Ltd. (Wuhan, China). Deionized water (18 MΩ cm) was used in all experiments.
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