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Live dead near infrared viability dye

Manufactured by Thermo Fisher Scientific
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The LIVE/DEAD near-infrared viability dye is a fluorescent reagent used to assess cell viability in a variety of cell-based applications. It utilizes near-infrared excitation and emission wavelengths to provide reliable and reproducible live/dead cell discrimination.

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Lab products found in correlation

Fugene 6 transfection reagent, by Promega (2 mentions) Anti mr1 pe antibody, by BioLegend (2 mentions) Cd45 clone 30 f11, by BD (1 mentions) Ly6g clone 1a8, by BD (1 mentions) Cd62l clone mel 14, by BD (1 mentions) Cd69 clone h1.2f3, by BD (1 mentions) Cd3 clone 17a2, by BioLegend (1 mentions) Cd11b clone m1 70, by BD (1 mentions) Cd4 clone gk1, by BioLegend (1 mentions)

Close spelling variants of this product

Live/Dead Near-InfraRed dye Near-infrared dye Live/Dead dye Viability dye

5 protocols using live dead near infrared viability dye

1

Immune Cell Profiling in Murine Super-Infection

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Whole blood was collected from mice on days −1, 3, 7 and 9 post‐super‐infection. Blood was collected in 0.5 M EDTA, then lysed using ACK Lysis buffer, washed and resuspended in flow cytometry buffer (2% FCS + 0.5 mM EDTA in PBS). Single‐cell suspensions were blocked with anti‐CD16/32 antibody and stained with fluorochrome‐conjugated antibodies against mouse CD3 (clone 17A2; Biolegend, San Diego, CA, USA), CD4 (clone GK1.5, Biolegend), CD8 (clone 53‐6.2, BD Biosciences, Franklin Lakes, NJ, USA), CD45 (clone 30‐F11, BD Biosciences), CD11b (clone M1/70, BD Biosciences), IA/IE (M5/114; eBioscience, San Diego, CA, USA), Ly6G (clone 1A8, BD Biosciences), CD62L (clone MEL‐14, BD Biosciences) and CD69 (clone H1.2F3, BD Biosciences), and dead cells were excluded using LIVE/DEAD Near‐Infrared viability dye (#L110909, Thermo Fisher Scientific). Sphero™ Blank Calibration Particles (BD Biosciences) were added to the samples and used as reference counting beads. Cells were acquired on a BD LSR II Fortessa Cell Analyser, and data were analysed using FlowJo software (v10.6; Treestar, Inc.).
Zaman M., Huber V.C., Heiden D.L., DeHaan K.N., Chandra S., Erickson D., Ozberk V., Pandey M., Bailly B., Martin G., Langshaw E.L., Zaid A., von Itzstein M, & Good M.F. (2021). Combinatorial liposomal peptide vaccine induces IgA and confers protection against influenza virus and bacterial super‐infection. Clinical & Translational Immunology, 10(9), e1337.
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2

Characterizing TCR Expression in HEK293T

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HEK293T cells were transiently co‐transfected with pMIG.TCR and pMIG.huCD3 plasmids using FuGENE 6 transfection reagent (Promega). Cells were cultured for 48–72 h, harvested by mechanical disruption and stained with anti‐CD3 (BV421, clone UCHT1; BD), LIVE/DEAD near‐infrared viability dye (Thermo Fisher Scientific) and MR1 tetramers. Cells were then fixed with 2% paraformaldehyde and analyzed by flow cytometry.
Seneviratna R., Redmond S.J., McWilliam H.E., Reantragoon R., Villadangos J.A., McCluskey J., Godfrey D.I, & Gherardin N.A. (2022). Differential antigenic requirements by diverse MR1‐restricted T cells. Immunology and Cell Biology, 100(2), 112-126.
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3

Transient TCR Expression in HEK293T

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HEK293T cells were transiently co-transfected with pMIG.TCR and pMIG.huCD3 plasmids using FuGENE 6 transfection reagent (Promega). Cells were cultured for 48–72 h, harvested by mechanical disruption and stained with anti-CD3 (BV421, clone UCHT1; BD), LIVE/DEAD near-infrared viability dye (Thermo Fisher Scientific) and MR1 tetramers. Cells were then fixed with 2% paraformaldehyde and analyzed by flow cytometry.
Seneviratna R., Redmond S.J., McWilliam H.E., Reantragoon R., Villadangos J.A., McCluskey J., Godfrey D.I, & Gherardin N.A. (2022). Differential antigenic requirements by diverse MR1‐restricted T cells. Immunology and Cell Biology, 100(2), 112-126.
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4

Quantifying Antigen-Induced MR1 Expression

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For antigen-induced MR1-upregulation assays, adherent APC lines were plated the day prior to the addition of antigen, whereas nonadherent lines were plated on the same day as the addition of antigen as above. APC lines were incubated with antigen for 4 h before being harvested. Adherent cell lines were mechanically disrupted. Cells were then stained with LIVE/DEAD near-infrared viability dye (Thermo Fisher Scientific) and anti-MR1-PE antibody (Clone 26.5, BioLegend), fixed in 2% paraformaldehyde and acquired by flow cytometry. For analysis, APCs were gated based on forward scatter-area and side scatter-area after dead cell and doublet removal.
Seneviratna R., Redmond S.J., McWilliam H.E., Reantragoon R., Villadangos J.A., McCluskey J., Godfrey D.I, & Gherardin N.A. (2022). Differential antigenic requirements by diverse MR1‐restricted T cells. Immunology and Cell Biology, 100(2), 112-126.
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5

Measuring MR1 Upregulation in APCs

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For antigen‐induced MR1‐upregulation assays, adherent APC lines were plated the day prior to the addition of antigen, whereas nonadherent lines were plated on the same day as the addition of antigen as above. APC lines were incubated with antigen for 4 h before being harvested. Adherent cell lines were mechanically disrupted. Cells were then stained with LIVE/DEAD near‐infrared viability dye (Thermo Fisher Scientific) and anti‐MR1‐PE antibody (Clone 26.5, BioLegend), fixed in 2% paraformaldehyde and acquired by flow cytometry. For analysis, APCs were gated based on forward scatter‐area and side scatter‐area after dead cell and doublet removal.
Seneviratna R., Redmond S.J., McWilliam H.E., Reantragoon R., Villadangos J.A., McCluskey J., Godfrey D.I, & Gherardin N.A. (2022). Differential antigenic requirements by diverse MR1‐restricted T cells. Immunology and Cell Biology, 100(2), 112-126.
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