In Situ Cell Death Detection Kit, TMR red

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Fix cell samples with Fixation solution for 1 h. Incubate in Permeabilisation solution (0.1% Triton X-100 in PBS) for 2 min on ice. Label and incubate for 60 min at +37°C in a humidified atmosphere in the dark	If low labelling is found, reduce fixation time or try 2% buffered paraformaldehyde

Protocol tips
Fix cell samples with Fixation solution for 1 h. Incubate in Permeabilisation solution (0.1% Triton X-100 in PBS) for 2 min on ice. Label and incubate for 60 min at +37°C in a humidified atmosphere in the dark
Downstream tips
If low labelling is found, reduce fixation time or try 2% buffered paraformaldehyde

Publication protocol

Cells were seeded on pre-coated coverslips (Nalge Nunc International, USA). Following treatment, the cells were fixed in 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min followed by permeabilization with 0.1% Triton X-100 in PBS for 5 min. After three washes with PBS, the specimens were labeled using an In Situ Cell Death Detection Kit (TMR red) (Roche Applied Science). Fluorescent images were observed using a DSU fluorescence microscope (BX 51; Olympus, Japan) coupled to a CCD camera (DP72, Olympus).

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for In Situ Cell Death Detection Kit, TMR red below.

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