Dead cell marker

Manufactured by Thermo Fisher Scientific

The Dead Cell Marker is a fluorescent dye that can be used to identify dead or dying cells in a sample. It binds to DNA and emits a fluorescent signal when excited by the appropriate wavelength of light. This allows researchers to distinguish between live and dead cells in their experiments.

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Lab products found in correlation

Perm2 permeabilizing solution, by BD (2 mentions) Facs lysing solution, by BD (2 mentions) Lsrfortessa, by BD (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

Spelling variants of this product

Live/dead fixable aqua dead cell marker (5 citations) LIVE/DEAD Fixable Near-IR Dead Cell Marker (2 citations) Live/Dead Aqua cell marker (2 citations)

3 protocols using dead cell marker

Multiparameter Flow Cytometric Analysis

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Freshly isolated PBMCs were stained with dead cell marker (Invitrogen, Carlsbad) and fluorochrome‐conjugated antibodies directed against intracellular and surface markers (Supplementary table 1) for 20 min at RT in the dark. Cells were washed two times with 150 µL of PBS with 10% FCS. Next, pellets were resuspended in 100 µL of BD FACS Lysing Solution (BD Biosciences) and incubated for 10 min at RT, for cell fixation. After washing with PBS with 10% FCS, cells were permeabilised using 100 µL of BD Perm2 Permeabilizing Solution (BD Biosciences) for 10 min at RT. Subsequently, antibodies were added for intracellular staining and incubated for 30 min at RT in the dark. Cells were then washed with 150 µL of PBS with 10% FCS and incubated in a 1% formaldehyde solution for 2 h, washed and resuspended in PBS containing 10% FCS.
Samples were acquired on a BD LSR Fortessa™. Flow cytometric analysis was performed using FlowJo version 10.6.2 (TreeStar, Ashland).

García M., Kokkinou E., Carrasco García A., Parrot T., Palma Medina L.M., Maleki K.T., Christ W., Varnaitė R., Filipovic I., Ljunggren H., Björkström N.K., Folkesson E., Rooyackers O., Eriksson L.I., Sönnerborg A., Aleman S., Strålin K., Gredmark‐Russ S., Klingström J, & Mjösberg J. (2020). Innate lymphoid cell composition associates with COVID‐19 disease severity. Clinical & Translational Immunology, 9(12), e1224.

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Detailed PBMC Immunophenotyping Protocol

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Freshly isolated PBMCs were stained with dead cell marker (Invitrogen) and fluorochrome-conjugated antibodies directed against intracellular and surface markers (See Table S1 in the Online Repository) for 20 min at RT in the dark. Cells were washed two times with 150 ul of PBS with 10% FCS. Next, pellets were resuspended in 100 ul of BD FACS Lysing Solution (BD Biosciences) and incubated for 10 min at RT, for cell fixation. After washing with PBS with 10% FCS, cells were permeabilized using 100 ul of BD Perm2 Permeabilizing Solution (BD Biosciences) for 10 min at RT. Subsequently, antibodies were added for intracellular staining and incubated for 30 min at RT in the dark. Cells were then washed with 150 ul of PBS with 10% FCS and incubated in a 1% formaldehyde solution for 2h, washed and resuspended in PBS containing 10% FCS.
The antibodies and dead cell marker used for flow cytometry staining are listed in Supplementary Table III. Samples were acquired on a BD LSR Fortessa TM . Flow cytometric analysis was performed using FlowJo version 10.6.2 (TreeStar).

García M., Kokkinou E., García A.C., Parrot T., Medina L.M.P., Maleki K.T., Christ W., Varnaitė R., Filipovic I., Ljunggren H., Björkström N.K., Folkesson E., Rooyackers O., Eriksson L.I., Sönnerborg A., Aleman S., Strålin K., Gredmark‐Russ S., Klingström J., & Mjösberg J. (2020). Innate lymphoid cell composition associates with COVID-19 disease severity.

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Multiparametric Flow Cytometry of Tumor and Lung Cells

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Isolated tumor-infiltrating cells and lung resident cells were stained with eFluor450-labeled anti-CD45.2, APC-labeled anti-CD11b, and FITC-labeled anti-Gr-1. All antibodies were purchased from eBioscience (San Diego, CA). Next, the cells were washed twice with PBS and then stained for dead cells with the Dead Cell Marker (Invitrogen, Grand Island, NY). Cells were acquired on a LSR-II flow cytometer (BD Bioscience) and thereafter analyzed using FlowJo software (Tree Star, Ashland, OR).

Weiss I.D., Ella E., Dominsky O., Smith Y., Abraham M., Wald H., Shlomai Z., Zamir G., Feigelson S.W., Shezen E., Bar-Shai A., Alon R., Izhar U., Peled A., Shapira O.M, & Wald O. (2015). In the hunt for therapeutic targets: mimicking the growth, metastasis, and stromal associations of early-stage lung cancer using a novel orthotopic animal model. Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, 10(1).

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