The cells were fixed and permeabilized as previously depicted.25 (link) Vinculin, the adhesion protein, was labelled with mouse monoclonal anti-vinculin primary antibody (1:200, Sigma, USA) and conjugated anti-Mouse IgG Secondary Antibody (1:1000, Invitrogen, USA). To label the F-actin structure, the samples were stained with phalloidin (Molecular Probes, USA) with cell nuclei visualized by incubation with l DAPI (5 μg/m, Dojindo, USA). Slides were then imaged with a fluorescence microscope (SP5, Leica, Germany) with x63 objective.
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Visualizing Cytoskeletal Structures
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Anti antibody
Anti igg
Antibody
Cell nuclei
Cells
Dapi
F actin
Fluorescence microscope
Molecular probes
Monoclonal antibody
Mouse
Phalloidin
Protein
Vinculin
Corresponding Organization :
Other organizations : XinHua Hospital, Shanghai Jiao Tong University, University of Nottingham
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Variable analysis
independent variables
- Fixation and permeabilization of cells as previously depicted
- Vinculin (adhesion protein) labeling
- F-actin structure labeling
- Cell nuclei visualization
- Mouse monoclonal anti-vinculin primary antibody (1:200, Sigma, USA)
- Conjugated anti-Mouse IgG Secondary Antibody (1:1000, Invitrogen, USA)
- Phalloidin (Molecular Probes, USA) for F-actin staining
- DAPI (5 μg/m, Dojindo, USA) for cell nuclei staining
- Fluorescence microscope (SP5, Leica, Germany) with x63 objective
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