LIVE/DEAD Fixable Violet Dead Cell Stain Kit

Protocol tips

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	Add 1 μL of the reconstituted fluorescent reactive dye and incubate at room temperature or on ice for 30 minutes, protected from light.

Protocol tips
Add 1 μL of the reconstituted fluorescent reactive dye and incubate at room temperature or on ice for 30 minutes, protected from light.

Publication protocol

ViViD, an amine reactive dye (LIVE/DEAD fixable violet dead cell stain; Thermo-Fisher Scientific, Loughborough, UK), was used to discriminate between live and dead cells. Stock solutions of LIVE/DEAD dye were prepared in dimethyl sulfoxide (DMSO; Sigma-Aldrich Company Ltd, Dorset, UK) according to the manufacturer's instructions. Cells were incubated with reconstituted LIVE/DEAD dye for 30 minutes in the dark. Media were removed, the cells resuspended in PBS, and then fixed with 4% paraformaldehyde for 15 minutes at room temperature. Fixative was replaced with 1 mL of PBS with 1% bovine serum albumin and the samples kept on ice. Samples were run on a LSRFortessa (BD Biosciences, Oxford, UK). Based on compensated green fluorescent protein (GFP)-fluorescence, gating settings were set to isolate GFP-positive, ViViD-positive, and negative cell fractions.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for LIVE/DEAD Fixable Violet Dead Cell Stain Kit below.

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