BmN-SWU1 cells were seeded at equal densities and cultured on coverslips in a 24-well plate. They were then treated with H2O2 as described above, washed in PBS and fixed in 4% PFA for 15 min. Then, they were permeabilized with 1% Triton X-100 in PBS for 15 min at room temperature and blocked with 1% BSA and 10% goat serum in PBS for 1 h at 37°C. The cells were then incubated with mouse anti-cytochrome c primary antibody (1:100; Beyotime) for 2 h and then stained with Cy3-labeled goat anti-mouse secondary antibody (1:500; Beyotime) for 1 h at 37°C. The nuclei were stained by incubation with DAPI for 8 min in the dark, and then mitochondria were stained with 300 nM Mito-Tracker Green solution for 20 min at 37°C. Finally, the cells were visualized through a confocal microscope.
Cy3 labeled goat anti mouse secondary antibody
Manufactured by Beyotime
Sourced in China
Cy3-labeled goat anti-mouse secondary antibody is a laboratory reagent used in various immunoassay techniques, such as Western blotting and immunofluorescence microscopy. It is composed of a goat-derived antibody specific to mouse immunoglobulins, conjugated with the Cy3 fluorescent dye. This secondary antibody is designed to detect and visualize the presence of primary mouse antibodies in experimental samples.
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Lab products found in correlation
2 protocols using cy3 labeled goat anti mouse secondary antibody
1
Cytochrome c Localization in Oxidative Stress
2
Immunofluorescence Analysis of Primary Trophoblasts
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In the present study, the primary trophoblast slides were first fixed with 4% paraformaldehyde for 15 min and then permeabilized with 0.1% Triton X-100 for 30 min at room temperature. Goat serum (purchased from Solarbio, Beijing, China) was added in order to block non-specific binding sites, followed by incubation with CK18 antibody (dilution 1:50; #sc-6259; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), vimentin antibody (dilution 1:100; #WL0274; Wanleibio, Shenyang, China) and hPL antibody (dilution 1:100; #ab11396; Abcam, Cambridge, MA, USA) at 4°C overnight. The slides were then washed three times with PBS for 5 min and incubated with Cy3-labeled goat anti-rabbit secondary antibody (#A0516) or Cy3-labeled goat anti-mouse secondary antibody (#A0521) (Beyotime Institute of Biotechnology, Haimen, China) for 1 h at room temperature. Subsequently, 4′,6-diamidino-2-phenylindole (DAPI) was added in order to stain nuclei, and they were then examined by fluorescence microscopy (BX53; purchased from Olympus, Tokyo, Japan).
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