Viability dye 7-AAD - (Jul/21/2014 )
Does anyone have experience with this dye 7-AAD? I am wondering when to add it to my cells ? i.e. prior to doing my cytofix/cytoperm or at the end of staining with everything.
I am doing a 10-color panel, which includes three intracellular cytokines (TNF, IFNg, IL-2: obviously needing cytofix/cytoperm)
7AAD works on primary live cells, and is not fixable.
You add it right before acquiring live cells on the cytometer. It can be used to stain already fixed, permabilized cells for DNA quantitation.
For fixable viability dyes, look at Invitrogen/Molecular Probes LIVE/DEAD series - available in a broad spectrum.
Similar fixable dyes are available from BioLegend and eBiosciences.
Hi. I have the same confusion as well. I wanted to use 7AAD in conjunction with intracellular staining
If stain the cells with 7AAD, then proceed to fixation and permeabilization for intracellular staining? Would that work?
7-AAD can be used with surface antigen staining/ antibodies but not intracellular markers from the datasheets I have been reading.