Duolink reagent

Manufactured by Merck Group

Duolink reagent is a laboratory product provided by Merck Group. It is a tool used for proximity ligation assay (PLA), a technique that allows the detection and visualization of protein-protein interactions within cells.

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2 protocols using duolink reagent

Proximity Ligation Assay for Flag-RAD50

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U2OS cells grown on coverslips were transfected with Flag-RAD50 or empty Flag-plasmid. Cells were permeabilized with 0.2% Triton X-100 for 5 minat room temperature and proximity ligation assay (PLA) was performed using Flag, MAP7 antibodies and Duolink reagent (Sigma-Aldrich) according to the manufacturer's protocol. For PLA quantification Z-stack images were taken for each condition with the LSM710 microscope at x63 objective lens. PLA signal was used to define the axis and 10 z-stack slices were acquired per image, stacks were pulled together, and merged with DAPI for the analysis.

Dullovi A., Ozgencil M., Rajvee V., Tse W.Y., Cutillas P.R., Martin S.A, & Hořejší Z. (2023). Microtubule-associated proteins MAP7 and MAP7D1 promote DNA double-strand break repair in the G1 cell cycle phase. iScience, 26(3), 106107.

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Investigating Histone H1 and BRCA1 Interactions

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U2OS cells grown on coverslips were transfected with siRNAs against histone H1 or control siRNAs and treated or not with HU for 2 h. Cells were then permeabilized with 0.2% Triton X-100 for 5 min at room temperature, and the PLA was performed using H1.2, H1.3, and BRCA1 antibodies and Duolink reagent (Sigma-Aldrich) according to the manufacturer’s protocol.

Ozgencil M., Dullovi A., Christiane Higos R.C., Hořejší Z, & Bellelli R. (2023). The linker histone H1–BRCA1 axis is a crucial mediator of replication fork stability. Life Science Alliance, 6(9), e202301933.

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