Cells cultured on confocal dishes were fixed with 4% paraformaldehyde for 20 min at RT, permeabilized and incubated with primary antibodies at 4 °C. For in situ proximity ligation assay, protein–protein interactions between YAP or TAZ and p52 were detected with secondary proximity probes (Anti-Rabbit Plus and Anti-Mouse Minus) according to the Duolink in situ Fluorescence User Guide (Sigma-Aldrich).
Duolink in situ fluorescence user guide
Manufactured by Merck Group
Duolink In Situ Fluorescence User Guide is a lab equipment product from Merck Group. It provides a step-by-step guide for the Duolink In Situ Fluorescence assay, which is a tool for detecting and visualizing protein-protein interactions in fixed cells or tissue samples.
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3 protocols using duolink in situ fluorescence user guide
1
Analyzing Protein-Protein Interactions
2
Protein-Protein Interaction Analysis in Cells
3
Protein-Protein Interaction Analysis in Cells
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Cells cultured on glass surfaces were fixed with 4% paraformaldehyde for 20 min at room temperature, permeabilized and incubated with primary antibodies at 4°C. For frozen sections, permeabilization and antibody incubations were performed under identical conditions as described above. All primary antibodies were used at a 1:200 dilution. For in situ PLA, protein-protein interactions between Snail/Slug and YAP/TAZ were detected with secondary proximity probes (Anti-Rabbit Plus and Anti-Mouse Minus) according to the Duolink In Situ Fluorescence User Guide (Sigma-Aldrich). Images were captured with a Leica confocal microscope. The antibody information was provided in the Supplementary Table 2 .
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