Cellular DNA Fragmentation ELISA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 2X10^4 cells in 100ml cell culture medium for 96-well microplates and allowed to adhere for 24h.		Samples can be measured within 5 min after adding the stop solution at a wavelength of 450 nm on a Bio-Rad microplate reader with 690 nm as reference.

Upstream tips
Seed 2X10^4 cells in 100ml cell culture medium for 96-well microplates and allowed to adhere for 24h.
Downstream tips
Samples can be measured within 5 min after adding the stop solution at a wavelength of 450 nm on a Bio-Rad microplate reader with 690 nm as reference.

Publication protocol

For the assessment of apoptosis-induced DNA fragmentation2x10^4cells in 100ml cell culture medium were plated per well in 96-well micro-plates and allowed to adhere for 24 h(378C, 5% CO2). The assay was performed following the instructions in the user’s manual (Cellular DNA Fragmentation ELISA kit; Roche Diagnostics, Basel Switzerland). The three different concentrations of NCT used for detection of DNA fragments were 1.1, 2.2, and 3.3 mM, each concentration in doublets, 200ml per well. The cells were incubated for3 and 6 h, respectively, at 378C in a humidified atmosphere. The samples were measured within 5 min after adding the stop solution at a wavelength of 450 nm on a Bio-Rad microplate reader with 690 nm as reference (Bio-Rad, Model 550; Bio-Rad Laboratories)

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