Apoptosis/ Necrosis Assay Kit (blue, green, red) (ab176749)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
• Use untreated cells as negative and treat cells for 24 h with 1 μM docetaxel, an anticancer taxane drug, to use as positive control for apoptosis induction

Upstream tips
• Use untreated cells as negative and treat cells for 24 h with 1 μM docetaxel, an anticancer taxane drug, to use as positive control for apoptosis induction

Publication protocol

Apoptosis was detected with annexin V (green fluorescence Excitation/Emission, Ex/Em = 490/525 nm)—an indicator that binds cell surface phosphatidylserine—using the Apoptosis/Necrosis Detection Kit (blue, green, red; ab176749; Abcam) and Cell-IQ MLF live imaging system. The detection kit was used according to manufacturer’s instructions while omitting the blue dye for live cells due to the lack of an appropriate excitation light source. Untreated cells were used as negative and cells treated for 24 h with 1 μM docetaxel, an anticancer taxane drug, were used as positive control for apoptosis induction. Cancer cell lines were irradiated with 4 Gy and apoptosis was monitored over several time points at 2 h, 4 h, and 24 h and 48 h. Fluorescent images were processed and merged after which fluorescence intensity was quantified using ImageJ and graphs were plotted using GraphPad Prism. Additionally, percentage of cells with apoptotic features was subjectively assessed and plotted.

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Manufacturer protocol

Download the product protocol from Abcam for Apoptosis/ Necrosis Assay Kit (blue, green, red) (ab176749) below.

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