For immunofluorescence, 1 × 104 cells in 100 μl of medium were placed in 96-well plates (6055300, PerkinElmer). Cells were fixed with 4% paraformaldehyde for 20 min, permeabilized, and blocked with 0.5% Triton X-100 (T8200, Solarbio) for 20 min. After incubation with Actin-Tracker Red-594 (C2205S, Beyotime) for 30 min, the cells were incubated further with 4′,6-diamidino-2-phenylindole (C0060, Solarbio) for 5 min at a 1:1000 dilution. Fluorescence images were captured using Opera Phenix Plus (PerkinElmer, HH14001000). Image analysis was performed with Harmony software (https://support.myharmony.com/en-cn/download ).
Actin tracker red 594
Manufactured by Beyotime
Sourced in China
Actin-Tracker Red-594 is a fluorescent probe designed to specifically label and visualize actin filaments in cells. It is a high-affinity, red-fluorescent dye that binds to actin with minimal perturbation of actin dynamics.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Beyotime (2 mentions)
Opera phenix plus, by PerkinElmer (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Mini67 1kt, by Merck Group (1 mentions)
Ab15580, by Abcam (1 mentions)
Coralite488 conjugated affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
4 protocols using actin tracker red 594
1
Fluorescent Actin Cytoskeleton Imaging
2
Actin-Based Cytoskeleton Visualization
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PASMCs were seeded in 24-well plates at a density of 50,000 cells per well and cultured for 24 h. Cytoskeleton was stained with Actin-Tracker Red-594 (Beyotime, China) according to manufacturer’s instruction. Fluorescence microscopy was used to capture images.
3
ADSC-Exosome Uptake by Schwann Cells
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ADSC-Exos were labeled with PKH67 (MINI67-1KT; Sigma-Aldrich) according to the manufacturer's instructions. Schwann cells were seeded in confocal dishes and coculture with PKH67 labeled ADSC-Exos. After 24 h, Schwann cells were fixed with 4% formaldehyde and stained with Actin-Tracker Red-594 (C2205S; Beyotime) and DAPI (C1005; Beyotime). Finally, cells were visualized using a laser scanning confocal microscope.
4
Immunofluorescence Analysis of Ki67 Expression
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The cultured cells were fixed with 4% paraformaldehyde at room temperature for 20 min, then permeabilized with 0.3% Triton X-100 for 15 min, blocked with 3% BSA for 30 min. After discarding the block solution, the samples were incubated with Ki67 (1:300, ab15580, abcam, United Kingdom) overnight, and then incubated with CoraLite 488-conjugated Affinipure Goat Anti-Rabbit IgG (H + L) (SA00013-2, proteintech, China) for 1.5 h. The nucleus was stained with DAPI (C1002, Beyotime, China) and the cytoskeleton was stained with Actin-Tracker Red 594 (F-Actin, C2205S, Beyotime, China). The number of the cell proliferation marker Ki67 was observed and recorded with a fluorescence inverted microscope (Nikon, Japan) at a wavelength of 550 nm.
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