For β-catenin staining, the treated cells were washed with PBS three times and fixed, then treated with 0.1% Triton X (Beyotime, China). Subsequently, a blocking buffer was added and the primary antibody was incubated overnight at 4°C. The next day, the treated cells were incubated with a secondary antibody, then treated with DAPI. Finally, the images were captured by a microscope.
Triton x
Manufactured by Beyotime
Sourced in China
Triton X is a non-ionic detergent commonly used as a laboratory reagent. It is a clear, colorless liquid that serves as a surfactant, capable of solubilizing and dispersing biological molecules in aqueous solutions.
Automatically generated - may contain errors
Lab products found in correlation
Ti2 u fluorescence microscope, by Nikon (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Anti e cadherin, by Proteintech (1 mentions)
Fluorescence microscope, by Zeiss (1 mentions)
Eclipse ti, by Nikon (1 mentions)
Cy3 conjugated secondary antibody, by Wuhan Servicebio Technology (1 mentions)
Paraformaldehyde (pfa), by Beyotime (1 mentions)
6 protocols using triton x
1
Immunostaining Protocol for β-Catenin
2
E-cadherin Immunofluorescence Visualization
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PC cells were seeded into confocal dishes and treated with DMSO and ST. Thereafter, the cells were washed with PBS, immobilized with 4% formaldehyde, and permeabilized with 0.2% Triton X (Beyotime Biotechnology, Suzhou, China). The cells were blocked with 5% BSA and stained with anti-E-cadherin (1:200; Cat No. 20874-1-AP, Proteintech) antibody overnight at 4℃. Thereafter, cells were washed with PBS and incubated with CY3 label-secondary antibody. Cell nuclei were stained with 4,6-diamino-2-phenylindole. Finally, cells were visualized using a Ti2-U fluorescence microscope (Nikon, Japan) at 562 nm.
3
p65 Nuclear Localization Assay
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For p65 staining, the treated cells were washed with PBS and treated with 0.1% Triton X (Beyotime, China). Subsequently, blocking buffer was added and the primary antibody was incubated overnight at 4°C. Then, cells were treated with secondary antibody and DAPI. Images were captured by a microscope.
4
Immunofluorescence Analysis of Chondrocyte Markers
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After harvesting, CHs were fixed by 4% formaldehyde for 15 min, incubated in 0.1% Triton X (Beyotime, China) for 15 min, and blocked by 2% BSA (Sigma, United States) for 1 h. Then, the cells were incubated with anti-pSOX9, anti-MRTF, anti-collagen Ⅰ and Ⅱ, anti-IL-6 and 10, and anti-vinculin overnight at 4 °C. Then, the cells were incubated with DyLight 488 or 647 goat anti-rabbit IgG antibody for 1 h in the dark. In addition, nuclei were counterstained with DAPI and the cytoskeleton was stained with phalloidin. All the antibodies were purchased from Abcam. The images were taken by a fluorescence microscope (Zeiss, Germany) and analyzed using ImageJ software (National Institutes of Health, United States).
5
Immunostaining of Glioma Cells
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Glioma cells were seeded into confocal dishes, immobilized with 4% formaldehyde, and permeabilized with 0.2% Triton X (Beyotime Biotechnology, Suzhou, China). Following blocking with 5% BSA, the cells were stained with the anti-E-cadherin (1:200), Slug (1:200), β-catenin (1:200), TSPO (1:200), MAPKAPK3 (1:200), and HUR (1:200) antibodies overnight at 4°C. After washing with PBS and incubating with CY3 or FITC-labeled second antibody, the cell nucleus was stained with a 4,6-diamino-2-phenylindole reagent and visualized using a Ti2-U fluorescence microscope (Nikon, Japan) or Axiotron 300 XYZ confocal microscopy (Zeiss, Germany).
6
Fluorescence Imaging of Organelles in Rat Chondrocytes
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Rat chondrocytes were seeded in a 24-well plate and cultured overnight. After being transfected with CellLight-GFP-BacMam-2.0 (2 μL / 10,000 cells) for 24 h to track mitochondria or lysosomes, the cells were treated with RSL3, 4OHT, ScpI2, DIDS, CCCP, CQ or colchicine. The cells were then fixed in 4% paraformaldehyde (Beyotime, Shanghai, China), permeabilized with 0.2% Triton-X (Beyotime, Shanghai, China), and blocked with 2% BSA (Sigma, MO, USA) for 0.5 h at room temperature. After overnight incubation with SCP2 antibody (1:100) at 4 °C, the cells were incubated with a Cy3-conjugated secondary antibody (1:100; Servicebio, Wuhan, China) at room temperature for 1 h followed by Hoechst 33258 (10 μg/mL) for 5 min. Images were acquired using a fluorescence microscope (Nikon, Eclipse-Ti) : fluorescence excitation/emission 550⁄570 nm for SCP2, 458⁄520 nm for GFP, and 352⁄461 nm for Hoechst.
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