Mouse anti–ZO-1 (33-9100) and rabbit ZO-2 (38-9100) antibodies were from GE Healthcare, mouse Myc-Tag (9B11) antibody was from Cell Signaling Technology, rabbit anti-GFP (Ab290) and rabbit anti–N-cadherin (Ab76057) antibodies were from Abcam, and rabbit anti–β-catenin (C2206) antibody was from Sigma-Aldrich. Antibodies were validated by recognizing bands of the predicted size on immunoblots and by cellular immunolocalization where previously reported. Species-specific secondary antibodies for immunofluorescence (Cy2, Cy3, and Cy5 conjugated) and immunoblots (IR-labeled 680 and 790/800 antibodies) were from Jackson ImmunoResearch. Rhodamine phalloidin and Alexa Fluor 647 phalloidin were from GE Healthcare. Halo ocln was labeled using Janelia Fluor 646 (Grimm et al., 2015 (link)) at 1:200 in cell culture media plus serum, followed by four 10-min washes to allow free dye to diffuse out. Cells were otherwise fixed and stained normally.
Rhodamine phalloidin
Manufactured by GE Healthcare
Rhodamine phalloidin is a fluorescent stain used in microscopy to visualize and detect actin filaments in cells. It binds specifically to F-actin, allowing for the imaging and analysis of the cellular cytoskeleton.
Automatically generated - may contain errors
2 protocols using rhodamine phalloidin
1
Immunostaining of Cell-Cell Junctions
2
Daam1 Antibody Generation and Characterization
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The following antibodies and reagents were used: mouse anti-Daam1 C monoclonal antibodies were raised against mouse Daam1 peptide a.a.1057-1068 following standard procedures. Cloned lines were selected by ELISA and Western blotting and anti-Daam1 C monoclonal antibodies were purified from identified cell lines using a protein A column. Additional antibodies and reagents: mouse anti-human Daam1 N (a.a. 1–111) monoclonal antibody (M05) (Abnova), mouse anti-Actin monoclonal antibody (Chemicon), mouse anti-Myc (9E10) (Santa Cruz Biotechnology), rat anti-TER119-PE and rat anti-CD71-FITC (BD-Pharmingen), anti-PECAM-1 (MEC13.3)(BD-Pharmingen), goat anti-mouse Ig Horseradish Peroxidase-linked (GE healthcare), goat anti-rat Ig Horseradish Peroxidase-linked (GE healthcare), Rhodamine-phalloidin, Texas Red-X phalloidin, BODIPY488-phalloidin, and 4’,6-diamidini-2-phenylindole, dihydrochloride (DAPI) (BD Bioscience).
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