FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Precoating slides with poly-L-lysine may enhance cell adherence. Do not allow the samples to dry out. All other FragELTM kit components, with the exception of the Mounting Media, should be kept on ice during usage then promptly returned to -20C	Dilute 5X TdT Equilibration Buffer 1:5 with water. Place slides in a humidified chamber and incubate at 37C for 1 - 1.5 hours after ading TdT Labeling Reaction Mixture

Upstream tips
Precoating slides with poly-L-lysine may enhance cell adherence. Do not allow the samples to dry out. All other FragELTM kit components, with the exception of the Mounting Media, should be kept on ice during usage then promptly returned to -20C
Protocol tips
Dilute 5X TdT Equilibration Buffer 1:5 with water. Place slides in a humidified chamber and incubate at 37C for 1 - 1.5 hours after ading TdT Labeling Reaction Mixture

Publication protocol

The MTT assays were performed after transfection for 0, 24, 48, and 96 hours as described previously (45).

Flow cytometric analysis was conducted at the Flow Cytometry facility of Peking Union Medical College. Briefly, after 48 to 72 hours of transfection, the cells were fixed and then incubated in PBS containing Rnase A (100 μg/ml; Sigma-Aldrich) and propidium iodide (50 μg/ml; Sigma-Aldrich) at room temperature for 30 minutes prior to analysis.

Glioma cells were infected with siRNAs or plasmids in 24-well plates for 48 to 72 hours. Then, the TUNEL assay was performed using a FragEL DNA fragmentation detection kit (Merck) according to the manufacturer’s instructions.

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Manufacturer protocol

Download the product protocol from Millipore for FragEL™ DNA Fragmentation Detection Kit, Colorimetric - TdT Enzyme below.

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Videos

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