The Duolink II fluorescence assay was used to analyze the interaction between YY1AP1 and YY1 in the HepG2-Tet-C2 cells. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100 for 10 minutes, then blocked and incubated with mouse anti-YY1AP1 Ab (1:100 dilution; Sigma) and rabbit anti-YY1 Ab (1:100 dilution; Abcam) overnight at 4°C. The Duolink II Proximity Ligation Assay (PLA) was then performed according to manufacturer's manual (Olink Bioscience, Uppsala, Sweden). A reporter substrate is formed if the PLA® probes are in very close proximity to one another indicating an interaction between the proteins examined. The reporter substrate appears as a red dot under a standard microscope. After mounting, the cells were visualized using a multiphoton confocal laser-scanning microscope (Carl Zeiss). Ten images per condition were acquired at 100× magnification with an average of ten cells per image. The number of red dots per cell was quantified from the average number of red dots per cell in ten images.
Duolink 2 proximity ligation assay
Manufactured by Olink
Sourced in Sweden
The Duolink II Proximity Ligation Assay is a laboratory tool that enables the detection and analysis of protein-protein interactions and modifications within cells. It is designed to amplify and detect these interactions through a signal-based approach.
Automatically generated - may contain errors
2 protocols using duolink 2 proximity ligation assay
1
Investigating Protein-Protein Interactions with Duolink II
2
Investigating Protein-Protein Interactions with Duolink II
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The Duolink II fluorescence assay was used to analyze the interaction between YY1AP1 and YY1 in the HepG2-Tet-C2 cells. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100 for 10 minutes, then blocked and incubated with mouse anti-YY1AP1 Ab (1:100 dilution; Sigma) and rabbit anti-YY1 Ab (1:100 dilution; Abcam) overnight at 4°C. The Duolink II Proximity Ligation Assay (PLA) was then performed according to manufacturer's manual (Olink Bioscience, Uppsala, Sweden). A reporter substrate is formed if the PLA® probes are in very close proximity to one another indicating an interaction between the proteins examined. The reporter substrate appears as a red dot under a standard microscope. After mounting, the cells were visualized using a multiphoton confocal laser-scanning microscope (Carl Zeiss). Ten images per condition were acquired at 100× magnification with an average of ten cells per image. The number of red dots per cell was quantified from the average number of red dots per cell in ten images.
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