Aqua amine fixable live dead dye

Manufactured by Thermo Fisher Scientific

Aqua Amine fixable live dead dye is a fluorescent stain used to distinguish between live and dead cells. It is designed to permeate cell membranes and bind to cellular proteins, allowing for the identification of viable and non-viable cells in a sample.

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Lab products found in correlation

Mouse tumor dissociation kit, by Miltenyi Biotec (2 mentions) Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions) Intracellular fixation permeabilization buffer set, by Thermo Fisher Scientific (1 mentions) Human truestain fcx, by BioLegend (1 mentions) Pd 1 pe, by BD (1 mentions) Truestain fcx, by BioLegend (1 mentions) B220 pecy5, by BioLegend (1 mentions) Cd3e pe cy5, by BioLegend (1 mentions) Xcr1 pe, by BD (1 mentions) Anti mouse cd80 apc, by BD (1 mentions) Cd4 alexa fluor 488, by BioLegend (1 mentions) Cd19 pe cy5, by BioLegend (1 mentions) Cd45 alexa fluor 700, by BioLegend (1 mentions)

Spelling variants of this product

Live/Dead Aqua (379 citations) LIVE/DEAD Fixable Aqua (214 citations) Live/dead fixable dye (56 citations) LIVE/DEAD Fixable Aqua dye (43 citations) Aqua Live/Dead dye (39 citations) LIVE/DEAD Aqua amine dye (7 citations) LIVE/DEAD Fixable Aqua Dead (7 citations) LIVE/DEAD aqua amine (6 citations) Fixable Aqua dye (4 citations) Aqua amine dye (3 citations)

2 protocols using aqua amine fixable live dead dye

Tumor Immune Cell Profiling

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Tumor tissues were digested using Mouse Tumor Dissociation kits (Miltenyi) according to the manufacturer’s protocols. Isolated cells were stained for viability using the Aqua Amine fixable live dead dye (Thermo Fisher) at room temperature using standard protocols. After viability staining, cells were resuspended in FACS buffer (PBS with 0.5% BSA and 2 mM EDTA) and Fc blocked using human TrueStain FcX (Biolegend). For cell surface staining, cells were incubated for 30 minutes at 4C with the following anti-mouse antibodies from Biolegend CD4-FITC, clone GK1.5, CD8a-BV786, clone 53–6.7; CD3e-Percp-Cy5.5, clone 17A2; and ThermoFisher CD45-AlexaFluor 700, clone 30-F11. Cells were then washed twice, fixed, and permeabilized (eBiosciences Intracellular Fixation & Permeabilization Buffer Set) prior to intracellular staining with FOXP3-PE, clone 150D. Samples were analyzed using an Attune NxT flow cytometer (Thermo Fisher) and data was analyzed using FlowJo 10 (TreeStar).

Knorr D., Leidner R., Jensen S., Meng R., Jones A., Ballesteros-Merino C., Bell R.B., Baez M., Sprott D., Bifulco C., Piening B., Dahan R., Fox B.A, & Ravetch J. (2023). FcyRIIB is a novel immune checkpoint in the tumor microenvironment limiting activity of Treg-targeting antibodies. bioRxiv.

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Comprehensive Immune Profiling of Tumor Cells

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Tumor tissues were digested using Mouse Tumor Dissociation kits (Miltenyi) according to the manufacturer’s protocols. Isolated cells were stained for viability using the Aqua Amine fixable live dead dye (Thermo Fisher) using standard protocols. After viability staining, cells were resuspended in FACS buffer (PBS with 0.5% BSA and 2 mM EDTA) and Fc blocked using mouse or human TrueStain FcX (Biolegend). For analysis stains, cells were incubated for 30 minutes at 4C with the following anti-mouse antibodies from Thermo Fisher: CD90.2 (Thy-1.2) SuperBright 645, clone 53-2.1; CD4-AlexaFluor 488, clone GK1.5, CD8a-SuperBright 780, clone 53-6.7; CD3e-PE-Cy5, clone 145-2C11; B220-PE-Cy5, clone RA3-6B2; CD19-PE-Cy5, clone 1D3; CD11c-PE-eFluor610, clone N418; CD45-AlexaFluor 700, clone 30-F11; MHCII-APC-eFluor 780, clone M5/114.15.2; antibodies from Biolegend: NK1.1-PE-Cy5, clone PK136; F4/80-AlexaFluor488; XCR-1-PE, clone ZET; anti-mouse CD80-APC, clone 16-10A1; CD86-Bv785, clone GL-1, PD-1-PE, clone 29F.1A12; LAG-3-PE-Cy7, clone C987W; anti-human CD40-Bv421, clone 5C3; antibodies from BD: anti-mouse CD40-Bv421, clone 3/23; anti-mouse CD172a-Bv605 (Sirpα), clone P84. Samples were analyzed using an Attune NxT flow cytometer (Thermo Fisher) and data was analyzed using FlowJo 10 (TreeStar).

Garris C.S., Wong J.L., Ravetch J.V, & Knorr D.A. (2021). Intravesical dendritic cell targeting with Fc-enhanced CD40 agonistic antibodies induces durable bladder cancer immunity. Science translational medicine, 13(594), eabd1346.

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