Cell Death Detection ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- Slides were fixed with 4% paraformaldehyde for 30 min at room temperature. - Slides were permeabilized with 1% Triton X-100 for 4 min at 4°C instead of 2min on ice. - Slides were labelled at 37°C for 60 min in a dark environment

Protocol tips
- Slides were fixed with 4% paraformaldehyde for 30 min at room temperature. - Slides were permeabilized with 1% Triton X-100 for 4 min at 4°C instead of 2min on ice. - Slides were labelled at 37°C for 60 min in a dark environment

Publication protocol

DNA fragmentation in PASMCs was measured using the TUNEL assay (Cell Death Detection Kit, Roche Biochemicals, Mannheim, Germany). Briefly, air-dried slides were fixed with 4% paraformaldehyde for 30 min at room temperature, cleaned three times with PBS for 10 min, and then permeabilized with 1% Triton X-100 for 4 min at 4°C. Then, the TdT-labelled nucleotide mix was added to each slide and incubated at 37°C for 60 min in a dark environment. The slides were washed twice with PBS and then counterstained with 10 mg/ml 4,6-diamidino-2-phenylindole (DAPI) for 5 min at 37°C [23].

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Manufacturer protocol

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